Amplification: ConsumablesAmplification Reagents and Plastics
Factors Impacting Gene Expression AnalysisRNA Isolation RNA integrity, purity, and yield■■ Genomic DNA contamination■■ Inhibitors of cDNA synthesis and qPCR■■ RNase and DNase contamination■■Reagents — Reverse Transcription cDNA synthesis efficiency■■ RNA protection■■ Input RNA capacity■■ Accurate representation of mRNA■■Reagents — Real-Time qPCR Detection sensitivity■■■■ Assay specificity■■ Inhibitors in sample■■ Reproducibility of thermal cycling conditions and instrument compatibilityPCR Plastic Consumables Instrument compatibility■■ Optimum performance■■ Automation friendly■■ Potential source of contamination and inhibition■■
RNA Isolation■■ Kits are designed and formulated to assist in the isolation of highly pure and intact RNA from different starting materials■■ RNA is compatible with a variety of downstream applications – Real-time qPCR – Northern blotting – Microarray analysis – cDNA library construction■■ DNase treatment ensures genomic DNA removal
RNA Isolation iScript™ RT-qPCR Sample Preparation Reagent PureZOL™ RNA Isolation Reagent ■■ Reagent stabilizes RNA and removes genomic DNA in less than 10 min ■■ Single-solution format permits recovery of RNA from small quantities of ■■ Suitable for adherent or suspension animal cells tissues or cells, making it ideally suited for gene expression studieswww.bio-rad.com/iscript ■■ RT-qPCR is directly enabled from cells without RNA purification when ■■ Efficient RNA purification from cultured cells, yeast, viruses, and bacteria, combined with an iScript reverse transcription kit and real-time supermix as well as plant and animal tissues ■■ Reagent allows multiplex real-time detection of up to 4 targets from as few ■■ PureZOL efficiently lyses cells and tissues, deproteinates RNA, and as 10 cells inactivates endogenous nucleases in a single step ■■ Ideal for rapid, high-throughput gene expression analysis ■■ Scalable starting sample amount For more information, request bulletin 5736. ■■ Convenient isolation of RNA, DNA, and protein from the same sample 104 Aurum™ Vacuum Manifold ■■ Vacuum-mediated nucleic acid purification platform ■■ Versatile manifold format adaptable for 96-well plate or up to 10 3 18 spin columns RFU ■■ Manifold ensures fast, high-quality sample preparation while maintaining the simplicity of vacuum processing 10 2 ■■ Unique vacuum regulator design allows for complete control of negative pressure 0 10 20 30 40 Cycles iScript RT-qPCR sample preparation reagent generates linear results over varying input cell amounts. HeLa cells (125, 25, 5, and 1 cells/µl) were treated and analyzed for GAPDH expression levels using iScript cDNA synthesis kit and iQ ™ SYBR ® Green supermix on the CFX96™ real-time PCR detection system. RFU, relative fluorescence units. 732-6890, 100 ml PureZOL RNA Isolation Reagent 170-8899, 5 x 10 ml 732-6470, 1 unit iScript RT-qPCR Sample Preparation Reagent Aurum Vacuum Manifold Amplification Reagents and Plastics 6 Visit us on the Web at www.bio-rad.com.
Reagents —Reverse Transcription■■ Formulated for efficient reverse transcription across a broad linear dynamic range■■ Potent RNase A inhibitors protect RNA during setup and reverse transcription■■ Flexible input RNA capacity to suit different experimental needs■■ Optimized for gene expression analysis using real-time qPCR
Reagents Two-Step Reverse Transcription Reagents iScript™ Advanced cDNA Synthesis Kit for RT-qPCR iScript Reverse Transcription Supermix ■■Increased qPCR data throughput and cost effectiveness from a single 20 µl for RT-qPCRwww.bio-rad.com/ reverse transcription (RT) reaction ■■ 1-tube format for simple and fast setup, and reduced pipetting variabilityiscript ■■ Superior sensitivity and broad linear dynamic range for RT (7.5 µg–100 fg) ■■ Liquid format at –20°C offers superior stability and eliminates freeze/thaw cycle ■■ 2-tube kit (5x iScript reaction mix and iScript reverse transcriptase) for ease ■■ Superior sensitivity and broad linear dynamic range for RT (1 µg–100 fg) of use and reduced reaction setup time ■■ Optimized blend of oligo(dT) and random primers ensures complete and ■■ Optimized blend of oligo(dT) and random primers ensures complete and unbiased RNA sequence representation unbiased RNA sequence representation ■■ RNase H+ MMLV reverse transcriptase (preblended with RNase inhibitor) ■■ RNase H+ MMLV reverse transcriptase (preblended with RNase inhibitor) delivers high sensitivity for real-time RT-qPCR and eliminates additional delivers high sensitivity for real-time RT-qPCR and eliminates additional RNase H+ step RNase H+ step ■■ Potent blend of RNaseA inhibitor protects RNA during setup and RT ■■ Potent blend of RNaseA inhibitor protects RNA during setup and RT ■■ Short 40 min protocol allows fast qPCR data generation ■■ Short 35 min protocol allows fast qPCR data generation For more information, request bulletin 6031. For more information, request bulletin 6125. 35 104 — 1 µg RNA Average Cq SD CV, % — 100 ng 103 104 100 ng 21.35 0.123 0.576 30 — 10 ng RFU — 1 ng 100 pg 31.56 0.147 0.465 102 — 100 pg 25 — 10 pg RFU RFU — 1 pg Cq 0 10 20 Cycles 30 40 103 20 103 102 15 –2 –1 0 1 2 3 4 0 10 20 30 40 10 20 30 40 log starting quantity Cycles Cycles iScript advanced cDNA synthesis kit for RT-qPCR provides iScript reverse transcription supermix for RT-qPCR efficiently Excellent data reproducibility. PGK-1 mRNA (~160 bp), a gene that superior sensitivity and a broad linear dynamic range for reverse reverse transcribes RNA over a broad linear dynamic range encodes a glycolytic enzyme, was quantified using iScript reverse transcription. Total RNA (7.5 µg–1 pg) from HeLa cells was reverse for reliable gene expression analysis data. Different amounts of transcription supermix for RT-qPCR both with 100 ng () and 100 pg () transcribed using the iScript advanced cDNA synthesis kit for HeLa cell RNA (amounts shown in inset) were reverse transcribed of input RNA. For each input RNA, 48 individual RT reactions were RT-qPCR in a 20 µl reaction. A tenfold dilution of generated cDNA and one-tenth of the resulting cDNA was used as a template to performed and one-tenth of the resulting cDNA was used in the qPCR was used as template to amplify α-tubulin in a 10 µl qPCR reaction amplify β-actin gene (~90 bp) in 20 µl qPCR reactions with iQ™ reaction with SsoFast™ probes supermix. The gene expression analysis with iQ™ SYBR® Green supermix on a CFX384™ real-time PCR SYBR® Green supermix. Standard curve R 2 = 0.999, efficiency = data show excellent reproducibility both with high and low levels of input detection system. Efficiency = 90.7%, R2 = 0.999, slope = –3.57. Cq, 99.7%, slope = –3.33. RFU, relative fluorescence units. target mRNA. The ~10 Cq difference for the 1,000-fold dilution of RNA quantification cycle; RFU, relative fluorescence units. (100 ng–100 pg) demonstrates good reverse transcription efficiencies across different input RNAs. Cq, quantification cycle; RFU, relative fluorescence units. Amplification Reagents and Plastics 10 Visit us on the Web at www.bio-rad.com.
Reagents One-Step RT-qPCR Reagents iScript™ One-Step RT-PCR Kit with SYBR® Green Benefits of iScript one-step kits: ■■ For use on a broad range of real-time PCR instruments ■■ Provide powerful combination of iScript RNase H+ reverse transcriptasewww.bio-rad.com/iscript ■■ Extremely sensitive detection (100 ng–1 pg) of input RNA and antibody-mediated hot-start iTaq™ DNA polymerase Are ideal for rapid, high-throughput gene expression analysis iScript One-Step RT-PCR Kit for Probes ■■ ■■ For use with all types of hybridization probes, including dual-labeled ■■ Perform cDNA synthesis and qPCR in 1 tube, minimizing handling and oligonucleotide probes, FRET probes, and molecular beacons contamination risk ■■ Extremely sensitive detection (1 µg–1 pg) of input RNA For more information, request bulletin 3066. PCR baseline-subtracted curve fit, RFU PCR baseline-subtracted curve fit, RFU 10,000 1,000 1,000 100 100 10 0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48 0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 Cycles Cycles iScript™ one-step RT-PCR kit with SYBR® Green provides high reproducibility and iScript one-step RT-PCR kit for probes delivers unparalleled results over an sensitivity across a broad range of concentrations. Reactions were performed in triplicate, extremely wide dynamic range. RNA (1 µg–100 fg) isolated from HeLa cells using along with no-template controls, using GAPDH primers and 100 ng–100 fg total HeLa RNA. the Aurum™ total RNA kit was reverse transcribed and amplified using primers to Reactions were carried out on the iCycler iQ® real-time detection system. Standard curve β-actin and a FAM-labeled detection probe. Each dilution was performed in triplicate r = 1.000, efficiency = 95%, slope = –3.47. RFU, relative fluorescence units. and RT-PCR was carried out on the iCycler iQ detection system. Standard curve r = 1.000, efficiency = 97.2%, slope = –3.39. RFU, relative fluorescence units. Ordering Information Catalog # Description $ Two-Step Reverse Transcription Reagents One-Step RT-qPCR Reagents 170-8842 iScript Advanced cDNA Synthesis Kit for RT-qPCR, 50 x 20 μl reactions 170-8892 iScript One-Step RT-PCR Kit with SYBR Green, 50 x 50 μl reactions 170-8843 iScript Advanced cDNA Synthesis Kit for RT-qPCR, 250 x 20 μl reactions 170-8893 iScript One-Step RT-PCR Kit with SYBR Green, 200 x 50 μl reactions 170-8890 iScript cDNA Synthesis Kit, 25 x 20 μl reactions 170-8894 iScript One-Step RT-PCR Kit for Probes, 50 x 50 μl reactions 170-8891 iScript cDNA Synthesis Kit, 100 x 20 μl reactions 170-8895 iScript One-Step RT-PCR Kit for Probes, 200 x 50 μl reactions 170-8840 iScript Reverse Transcription Supermix for RT-qPCR, 25 x 20 μl reactions 170-8841 iScript Reverse Transcription Supermix for RT-qPCR, 100 x 20 μl reactions 170-8896 iScript Select cDNA Synthesis Kit, 25 x 20 μl reactions 170-8897 iScript Select cDNA Synthesis Kit, 100 x 20 μl reactions Amplification Reagents and Plastics 12 Visit us on the Web at www.bio-rad.com.
Reagents —Real-Time qPCR■■ Patented Sso7d fusion enzyme technology delivers higher processivity and inhibitor tolerance■■ Antibody-mediated hot-start polymerases enable instant activation and higher specificity■■ Choice of fast, standard, or universal cycling conditions■■ Formulated for optimal performance on a variety of real-time instruments Reagents
Reagents Real-Time qPCR Reagents Selection Guide SYBR® Green / EvaGreen Supermixes Probes Supermixes One-Step Kits for RT-qPCR SsoAdvanced™ SsoFast™ iQ™ SsoFast iTaq™ Fast iTaq™ EpiQ™ SsoFast iQ iQ SsoFast iTaq Fast iTaq iScript™ iScript SYBR® EvaGreen® SYBR® EvaGreen SYBR® Green SYBR® Chromatin Probes Supermix Multiplex Probes Supermix Supermix One-Step One-Step Green Supermix Green Supermix Supermix with Green SYBR® Supermix Powermix Supermix with ROX with ROX RT-PCR Kit RT-PCR Kit Supermix Supermix with Low ROX Supermix Green with ROX with SYBR® for ProbesReal-Time PCR Instrument ROX with ROX Supermix GreenBio-RadCFX96™, CFX96 Touch™,CFX384™, CFX384 Touch™, CFX ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔Connect™iQ™, iQ™5, MyiQ™, MyiQ™2 ✔ ✔ ✔ ✔ ✔ ✔■ ✔ ✔ ✔MiniOpticon™, DNA EngineOpticon® I and II ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔Applied BiosystemsStepOne/StepOne Plus ◆ ◆ ◆ ◆ ◆ ✔ ✔ ◆ ◆ ◆ ✔ ◆ ✔ ◆ ◆7500, ViiA7 ✔ ✔ ✔ ✔7000, 7300, 7700, 7900HT ✔ ✔ ✔StratageneMx3000P, 3005P, 4000 ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔EppendorfMastercycler eprealplex 2 or 4 ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔QIAGEN/CorbettRotor-Gene 3000, 6000, Q ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔RocheLightCycler 480 ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔LightCycler 1.0, 1.5, 2.0 ▲ ▲ ▲ ▲ ▲ ▲ ✔ ▲ ▲ ▲ ▲ ▲ ▲ ▲ ▲Idaho TechnologyLightScanner HR-1 ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔ ✔LightScanner 32 ▲ ▲ ▲ ▲ ▲ ▲ ✔ ▲ ▲ ▲ ▲ ▲ ▲ ▲ ▲✔ Recommended for use as is ◆ ROX reference setting must be turned “off” ▲ BSA must be added according to instrument specifications Amplification Reagents and Plastics 14 Visit us on the Web at www.bio-rad.com.
Reagents Real-Time qPCR — SYBR® Green/EvaGreen SsoFast™ EvaGreen® Supermix SsoFast EvaGreen Supermix with Low ROX ■■ Inhibitor tolerance and enhanced processivity with Sso7d fusion polymerase ■■ Blended with ROX for optimal performance on Applied Biosystemswww.bio-rad.com/ standard and fast 7500 real-time PCR instruments ■■ Robust formulation ensures maximum efficiency, sensitivity,supermixes and reproducibility for qPCR For more information, request bulletin 5919. ■■ Instant polymerase activation and rapid polymerization kinetics deliver fast qPCR results For more information, request bulletin 5816. 104 100.000 6,000 900 800 5,000 700 10.000 600 500 RFU 4,000 400 103 RFU RFU ∆Rn 300 3,000 1.000 200 100 2,000 0 0 10 20 30 40 0.100 Cycles 1,000 102 0 0.010 0 10 20 30 40 0 10 20 30 40 4 8 12 16 20 24 28 32 36 40 Cycles Cycles Cycles The unique fusion polymerase in SsoFast EvaGreen supermix SsoFast EvaGreen supermix demonstrates superior inhibitor The unique fusion polymerase in SsoFast EvaGreen supermix delivers extreme speed and generates exceptional qPCR results tolerance and direct qPCR capability from cell culture. Results with low ROX generates exceptional qPCR results on the ABI in less than 30 min. Tenfold serial dilutions of 10 ng–100 ag cDNA show efficient amplification and detection of a spike-in control 7500 fast real-time PCR system. Tenfold serial dilutions of 10 ng–100 fg from human spleen were used in each 20 μl reaction to detect template in the absence (■) or presence (■) of conditioned tissue cDNA from human spleen were used in each 20 μl reaction to detect 18S rRNA. 18S rRNA efficiency = 101.8%, r = 0.997. Total qPCR run culture medium using SsoFast EvaGreen supermix. Inset shows a α-tubulin. α-tubulin efficiency = 105.8%, r = 0.996. Total qPCR run time = 29 min. RFU, relative fluorescence units. competitor’s standard qPCR reagent is able to amplify the spike-in time = 39 min (not including melt curve). control template only in the absence (■) vs. presence (■) of culture medium. RFU, relative fluorescence units. Amplification Reagents and Plastics 16 Visit us on the Web at www.bio-rad.com.
Reagents Real-Time qPCR — SYBR® Green/EvaGreen Precision Melt Supermix EpiQ™ Chromatin SYBR® Green Supermix ■■ Optimized formulation containing EvaGreen dye delivers robust PCR and ■■ Robust formulation delivers superior sensitivity and efficiency for qPCR fromwww.bio-rad.com/ high resolution melt (HRM) performance genomic DNA templatessupermixes ■■ Sensitive and effective discrimination of all 4 SNP classes across a broad ■■ Protocol is optimized for difficult real-time qPCR reactions for high GC amplicons range of amplicons ■■ Supermix contains fluorescein and ROX and is compatible with all real- ■■ Accurate detection of CpG methylation status for epigenetic studies time PCR instruments except Applied Biosystems 7000, 7300, 7700, and 7900 models (additional ROX can be added by ordering the dye ■■ Exceptional room temperature stability for high-throughput HRM studies separately, catalog #172-5858) ■■ Reliable performance on any HRM-capable thermal cycler Please refer to page 22 for more details about the EpiQ chromatin analysis kit. For more information, request bulletin 6137. For more information, request bulletin 6020. A B 0.20 0.02 0.0 –0.1 0.15 0.00 Difference RFU –0.2 Difference RFU Difference RFU 0.10 –0.02 –0.3 –0.04 0.05 –0.4 –0.06 –0.5 0.00 –0.08 –0.6 –0.05 78 79 80 81 82 83 75 76 77 78 79 80 74 76 78 80 82 84 Temperature, °C Temperature, °C Temperature, °C Precision melt supermix delivers robust HRM for single nucleotide polymorphisms (SNPs). Discrimination of class I Accurate methylation detection with precision melt supermix. Mixtures and IV SNP genotypes are shown in panels A and B, respectively. Class I (A to G substitution) and class IV (A to T substitution) SNP of methylated and unmethylated human genomic DNA of varying ratios were genotypes from mouse genomic DNA were analyzed using precision melt supermix. Wild type (■), heterozygote (■), and homozygous analyzed using HRM on a CFX384 real-time PCR detection system. Increasing mutant (■) are shown in the difference plots normalized to wild-type samples. HRM analysis was performed on a CFX384™ real-time PCR amounts of methylated DNA (■, 0%; ■, 2%; ■, 5%; ■, 50%; ■, 75%; ■, 95%; detection system and genotypes were automatically assigned by Precision Melt Analysis™ software. Amplification was carried out for 35 and ■, 100%) were analyzed for methylation of the human RARB2 gene. The cycles. Total run time including melt curve = 150 min. RFU, relative fluorescence units. genomic region contains 7 CpG sites and is 88 base pairs in length. Total run time including melt curve = 190 min. RFU, relative fluorescence units. Amplification Reagents and Plastics 18 Visit us on the Web at www.bio-rad.com.
Reagents Real-Time qPCR Probes iQ™ Multiplex Powermix iTaq™ Fast Supermix with ROX ■■ Robust supermix formulated for sensitive and efficient multiplex qPCR ■■ Developed and validated for Applied Biosystems 7500 standard and fastwww.bio-rad.com/ and Stratagene real-time PCR instrumentssupermixes ■■ Reliable quantification of up to 4 targets (expression levels can vary up to 106-fold between target genes) or up to 5 targets ■■ Compatible with standard and fast thermal cycling conditions ■■ Linearity over 6 orders of magnitude of input cDNA and 4 orders of ■■ Antibody-mediated iTaq DNA polymerase provided in a convenient magnitude of input genomic DNA 2x supermix ■■ Suitable for a wide variety of applications, including gene expression For more information, request bulletin 5737. analysis, SNP genotyping, SNP analysis, GMO detection, and viral load detection iTaq Supermix with ROX ■■ Developed and validated for Applied Biosystems 7000, 7300, 7700, 7900, For more information, request bulletin 5348. StepOne, and StepOne Plus real-time PCR instruments ■■ Sensitive and accurate detection over 6 orders of magnitude ■■ Hot-start iTaq DNA polymerase in optimized buffer for simplex and duplex qPCR For more information, request bulletin 3065. 10.000 PCR baseline-subtracted curve fit, RFU 1,000 1.000 ∆Rn ∆Rn 1.000 100 0.100 0.100 0 5 10 15 20 25 30 35 40 45 0 5 10 15 20 25 30 35 40 45 0 10 20 30 40 Cycles Cycles Cycles iQ multiplex powermix produces highly reliable qPCR results Robust duplex qPCR results with iTaq fast supermix with ROX on the iTaq supermix with ROX produces superior results on the ABI 7900HT for up to five targets in a single tube, with no difference in Applied Biosystems 7500 fast real-time PCR system. cDNA inputs: system. Total RNA from HeLa cells was reverse transcribed using the detection of a low-expressing gene in multiplex or singleplex. 100-fold serial dilutions of cDNA from the equivalent of 1 µg–1 pg total RNA iScript™ reverse transcription supermix for RT-qPCR. A tenfold dilution of One-tenth of a 1 µg cDNA synthesis reaction of human thymus total were used in each 20 µl reaction. FAM-labeled 18S rRNA probe duplex generated cDNA (100 ng–1 pg) was used as template to amplify the β-actin RNA was used in each 20 µl reaction. FAM-labeled β-actin probe (), reaction (), VIC-labeled beta-2-microglobulin (B2M) probe duplex gene with a FAM-labeled probe. Standard curve R2 = 0.999, efficiency = Cy5-labeled α-tubulin probe (), HEX-labeled GAPDH probe (), reaction (), VIC-labeled B2M probe singleplex reaction (). 18S rRNA 98.0%, slope = –3.38. TAMRA-labeled cyclophilin probe (), Texas Red–labeled IL-2 probe (). efficiency = 98.6%, r = 0.999; B2M efficiency = 98.0%, r = 0.998. Total RFU, relative fluorescence units. qPCR run time = 45 min. Amplification Reagents and Plastics 20 Visit us on the Web at www.bio-rad.com.
Reagents Ordering Information Ordering Information Catalog # Description $ qPCR Supermixes with SYBR Green or EvaGreen qPCR Supermixes for Probes 172-5260 SsoAdvanced SYBR Green Supermix, 200 x 20 µl reactions 172-5230 SsoFast Probes Supermix, 200 x 20 µl reactions 172-5261 SsoAdvanced SYBR Green Supermix, 500 x 20 µl reactions 172-5231 SsoFast Probes Supermix, 500 x 20 µl reactions 172-5262 SsoAdvanced SYBR Green Supermix, 1,000 x 20 µl reactions 172-5232 SsoFast Probes Supermix, 1,000 x 20 µl reactions 172-5264 SsoAdvanced SYBR Green Supermix, 2,500 x 20 µl reactions 172-5233 SsoFast Probes Supermix, 2,000 x 20 µl reactions 172-5265 SsoAdvanced SYBR Green Supermix, 5,000 x 20 µl reactions 172-5250 SsoFast Probes Supermix with ROX, 200 x 20 µl reactions 172-5200 SsoFast EvaGreen Supermix, 200 x 20 µl reactions 172-5251 SsoFast Probes Supermix with ROX, 500 x 20 µl reactions 172-5201 SsoFast EvaGreen Supermix, 500 x 20 µl reactions 172-5252 SsoFast Probes Supermix with ROX, 1,000 x 20 µl reactions 172-5202 SsoFast EvaGreen Supermix, 1,000 x 20 µl reactions 172-5253 SsoFast Probes Supermix with ROX, 2,000 x 20 µl reactions 172-5203 SsoFast EvaGreen Supermix, 2,000 x 20 µl reactions 170-8860 iQ Supermix, 100 x 50 µl reactions 172-5204 SsoFast EvaGreen Supermix, 2,500 x 20 µl reactions 170-8862 iQ Supermix, 500 x 50 µl reactions 172-5205 SsoFast EvaGreen Supermix, 5,000 x 20 µl reactions 170-8864 iQ Supermix, 1,000 x 50 µl reactions 172-5210 SsoFast EvaGreen Supermix with Low ROX, 200 x 20 µl reactions 172-5848 iQ Multiplex Powermix, 50 x 50 µl reactions 172-5211 SsoFast EvaGreen Supermix with Low ROX, 500 x 20 µl reactions 172-5849 iQ Multiplex Powermix, 200 x 50 µl reactions 172-5212 SsoFast EvaGreen Supermix with Low ROX, 1,000 x 20 µl reactions 172-5105 iTaq Fast Supermix with ROX, 200 x 20 µl reactions 172-5213 SsoFast EvaGreen Supermix with Low ROX, 2,000 x 20 µl reactions 172-5106 iTaq Fast Supermix with ROX, 500 x 20 µl reactions 170-8880 iQ SYBR Green Supermix, 100 x 50 µl reactions 172-5107 iTaq Fast Supermix with ROX, 1,000 x 20 µl reactions 170-8882 iQ SYBR Green Supermix, 500 x 50 µl reactions 172-5108 iTaq Fast Supermix with ROX, 2,000 x 20 µl reactions 170-8884 iQ SYBR Green Supermix, 1,000 x 50 µl reactions 172-5854 iTaq Supermix with ROX, 200 x 50 µl reactions 170-8885 iQ SYBR Green Supermix, 2,000 x 50 µl reactions 172-5855 iTaq Supermix with ROX, 500 x 50 µl reactions 170-8886 iQ SYBR Green Supermix, 2,500 x 50 µl reactions 172-5856 iTaq Supermix with ROX, 1,000 x 50 µl reactions 170-8887 iQ SYBR Green Supermix, 5,000 x 50 µl reactions 172-5857 iTaq Supermix with ROX, 2,000 x 50 µl reactions 172-5100 iTaq Fast SYBR Green Supermix with ROX, 200 x 20 µl reactions Standard PCR Supermixes 172-5101 iTaq Fast SYBR Green Supermix with ROX, 500 x 20 µl reactions 172-5300 iProof High-Fidelity DNA Polymerase, 2 U/µl, 20 U 172-5102 iTaq Fast SYBR Green Supermix with ROX, 1,000 x 20 µl reactions 172-5301 iProof High-Fidelity DNA Polymerase, 2 U/µl, 100 U 172-5103 iTaq Fast SYBR Green Supermix with ROX, 2,000 x 20 µl reactions 172-5302 iProof High-Fidelity DNA Polymerase, 2 U/µl, 500 U 172-5850 iTaq SYBR Green Supermix with ROX, 200 x 50 µl reactions 172-5310 iProof HF Master Mix, 0.04 U/µl, 100 x 50 µl reactions 172-5851 iTaq SYBR Green Supermix with ROX, 500 x 50 µl reactions 172-5311 iProof HF Master Mix, 0.04 U/µl, 500 x 50 µl reactions 172-5852 iTaq SYBR Green Supermix with ROX, 1,000 x 50 µl reactions 172-5320 iProof GC Master Mix, 0.04 U/µl, 100 x 50 µl reactions 172-5853 iTaq SYBR Green Supermix with ROX, 2,000 x 50 µl reactions 172-5321 iProof GC Master Mix, 0.04 U/µl, 500 x 50 µl reactions 172-5110 Precision Melt Supermix, 200 x 20 µl reactions 170-8870 iTaq DNA Polymerase, 5 U/µl, 250 U 172-5112 Precision Melt Supermix, 1,000 x 20 µl reactions 170-8875 iTaq DNA Polymerase, 5 U/µl, 5,000 U 172-5404 EpiQ Chromatin SYBR Green Supermix, 500 x 20 µl reactions 170-8874 dNTP Mix, 200 µl 172-5405 EpiQ Chromatin SYBR Green Supermix, 1,000 x 20 µl reactions 172-5858 ROX Passive Reference Dye, 0.5 mlAmplification Reagents and Plastics 22 Visit us on the Web at www.bio-rad.com.
PCR Plastic Consumables■■ Precisely manufactured for optimal fit and cycling performance■■ Produced in Class 10,000 or 100,000 cleanroom environment■■ Certified to be free of DNase, RNase, and human genomic DNA■■ Extremely uniform wells reduce well-to-well variability in real-time PCR■■ Warp-free Hard-Shell® plates are designed for optimum performance with automation
PCR Plastic Consumables Capping Tools, Racks, and Multiplate™ Plates Capping Tools and Racks Multiplate 48-Well PCR Plates The versatile, unskirted design and 48-well format make these Multiplatewww.bio-rad.com/ unskirted PCR plates ideal for laboratories using 48-well blocks on Bio-Radpcrplastics instruments. The plates are suitable for reaction volumes of 5–125 µl. The polypropylene construction of Multiplate PCR plates confers very low protein binding and excellent preservation of sample volume. When less than a full plate is needed, these plates can be easily cut with scissors to the required size. Two plate styles are available: ■■ High-profile (20.70 mm) wells, clear — designed to fit in most thermal cyclers ■■ Low-profile (15.50 mm) wells, clear or white — optimized for fast PCR and TRC-9601, ANSI/SBS standard PCR Tube Rack low-volume reactions Compatible Instruments Bio-Rad C1000™, C1000 Touch™, T100™ (high-profile plates only), S1000™, DNA Engine® family, MJ Mini™, MiniOpticon™ (low-profile white plates recommended) ECT-2000 Strip Cap Tool MLP-4801, clear Multiplate 48-Well Unskirted PCR Plates TRC-0501, with covers 96-Place Racks ECT-1000 MLL-4801, clear Easy Cap™ Tool MLL-4851, white Multiplate Low-Profile 48-Well Unskirted PCR Plates Amplification Reagents and Plastics 28 Visit us on the Web at www.bio-rad.com.
PCR Plastic Consumables Hard-Shell® and Multiplate™ Plates Hard-Shell High-Profile 96-Well Semi-Skirted Multiplate 96-Well Unskirted PCR Plates PCR Plates The single-component polypropylene construction of Multiplate PCR plateswww.bio-rad.com/ confers very low protein binding and excellent retention of sample. When less These Hard-Shell PCR plates fit most thermal cyclers. Features of semi-pcrplastics skirted plates include: than a full plate is needed, these plates are easily cut with scissors to the required size. Two plate styles are available. ■■ Reaction volumes of 5–125 µl (350 µl maximum) High-profile for most standard instruments ■■ High-profile (20.75 mm) wells that fit most thermal cyclers, real-time PCR detection systems, and DNA sequencers Compatible Instruments ■■ Warp-free half-height skirt for improved robotic handling and labeling surface Bio-Rad C1000, C1000 Touch, S1000, T100, DNA Engine family, PTC-100, MyCycler™, ■■ Black alphanumeric labeling for easy well identification iCycler, MJ Mini™, iQ5, iCycler iQ, MyiQ, MyiQ2, Chromo4, MiniOpticon™; Applied Biosystems Compatible Instruments 0.2 ml tube cyclers, real-time systems, and Bio-Rad C1000™, C1000 Touch™, S1000™, T100™, DNA Engine® family, PTC-100®, iCycler ®, iQ™5, DNA sequencers; Eppendorf Mastercycler iCycler iQ®, MyiQ™, MyiQ™2, Chromo4™; Applied Biosystems 0.2 ml tube cyclers, real-time series; Stratagene (Agilent) Mx series; Idaho systems, and DNA sequencers; Eppendorf Mastercycler series; Stratagene (Agilent) Mx series; Technology LightScanner MLP-9601, clear Idaho Technology LightScanner MLP-9651, white Multiplate 96-Well Unskirted PCR Plates Low-profile for most fast instruments Compatible Instruments Bio-Rad C1000, C1000 Touch, S1000, DNA Engine family, PTC-100, MJ Mini, CFX96™, CFX96 Touch™, CFX Connect™, Chromo4, DNA Engine Opticon®; Applied Biosystems 0.1 ml tube fast cyclers and real-time systems; Eppendorf Mastercycler series; Idaho Technology LightScanner MLL-9601, clear MLL-9651, white Multiplate Low-Profile 96-Well Unskirted PCR Plates HSS-9601, clear shell, clear well HSS-9641, green shell, clear well HSS-9901, clear shell, clear well, bar-coded HSS-9665, black shell, white well Hard-Shell High-Profile 96-Well Semi-Skirted PCR Plates Amplification Reagents and Plastics 30 Visit us on the Web at www.bio-rad.com.
PCR Plastic Consumables Microseal® Sealers Microseal C Optical Seals Microseal F Foil Microseal C optical seals are optically clear adhesive films designed for These aluminized foil seals act as a barrier against evaporation from –80°C towww.bio-rad.com/ optical assays, such as those performed in real-time PCR. The seals can be 105°C. They are thin enough to pierce with a pipet tip for recovery of samplespcrplastics used for microplates with raised well rims. Features include: from individual wells, and they are suitable for use with automated systems, such as the ABI 3700 DNA analyzer. In addition to cold storage applications, ■■ Form a tight seal for PCR and qPCR they can be used for thermal cycling for sample volumes of 25 µl (96-well) or ■■ Pressure- and heat-sensitive adhesive sticks to the plate, not your gloves 5 µl (384-well). ■■ Sealing tool for applying even pressure is included ■■ Free from DNase, RNase, and human DNA contaminants Microseal A Film Microseal A film quickly and effectively seals the full range of Bio-Rad PCR Microseal B Adhesive Seals, Optically Clear plates and tubes. The pliant inner layer is designed to seal tightly during cycling Microseal B seals provide an adhesive-based sealing option for thermal yet to release smoothly to minimize the risk of aerosol formation and cross- cycling using thin-wall PCR plates. The strong adhesive layer ensures secure contamination of samples. This film is easily cut for use with fewer than 96 sample storage (from as low as –40°C and up to 110°C) or transport before or wells; a peel-away release liner protects the sealing surface from contamination after cycling, as well as tight sealing during thermal cycling when supplemental (not suitable for qPCR). An optional sealing roller provides a quick way to pressure is applied by a heated lid. This clear polyester film allows easy firmly seal Microseal A film on an entire array of wells. inspection of sample wells and effective light transmission for optical assays. The sealing surface is protected from contamination by a peel-away release liner. Perforated end-tabs allow removal of overhanging film for automation and other applications. Accessories include: ■■ Optical compression pad (96-well) — enhances the seal integrity of Microseal B clear seals when used in real-time PCR detection systems ■■ Optical film sealing kit — contains 100 Microseal B clear seals and an optical compression pad MSF-1001 MSA-5001 Microseal F Foil Microseal A Film MSC-1001 MSR-0001 MSB-1001 ADR-3296 Microseal C Optical Seals Sealing Roller Microseal B Adhesive Seals Optical Compression Pad Amplification Reagents and Plastics 32 Visit us on the Web at www.bio-rad.com.
PCR Plastic Consumables Ordering Information Ordering Information Description Clear Wells White Wells Black Wells Hard-Shell Low-Profile 96-Well Skirted PCR Plates White shell, 50 HSP-9601 HSP-9655 — Red shell, 50 HSP-9611 — — Yellow shell, 50 HSP-9621 — — Blue shell, 50 HSP-9631 HSP-9635 — Green shell, 50 HSP-9641 HSP-9645 — Black shell, 50 HSP-9661 HSP-9665 HSP-9666 White shell, bar-coded, 50 HSP-9901 HSP-9955 — Hard-Shell High-Profile 96-Well Semi-Skirted PCR Plates Clear shell, 25 HSS-9601 — — Green shell, 25 HSS-9641 — — Black shell, 25 — HSS-9665 — Clear shell, bar-coded, 25 HSS-9901 — — Hard-Shell 384-Well Standard PCR Plates Clear shell, 50 HSP-3801 HSP-3805 — White shell, 50 HSP-3851 — — Red shell, 50 HSP-3811 — — Yellow shell, 50 HSP-3821 — — Blue shell, 50 HSP-3831 — — Green shell, 50 HSP-3841 — — Black shell, 50 — HSP-3865 HSP-3866 Clear shell, bar-coded, 50 HSP-3901 HSP-3905 — 50 Plates 100 Plates, 100 Microseal C Seals Hard-Shell 384-Well 480 PCR Plates with Bar Code on Row A Side Clear shell, white well HSR-4805 HSR-4805K Clear shell, clear well HSR-4801 HSR-4801K Catalog # Description PCR Plate Sealers MSA-5001 Microseal A Film, package of 50 seals MSB-1001 Microseal B Adhesive Seals, optically clear, 100 seals MSC-1001 Microseal C Optical Seals, 100 seals MSF-1001 Microseal F Foil, package of 100 seals MSR-0001 Sealing Roller, for film seals ADR-3296 Optical Compression Pad, for improved film sealing of 96-well plates in DNA Engine Opticon 2 and Chromo4 systems ADR-5001 Pressure Pad, uniformly distributes lid pressure for sealing film MSO-1001 Optical Film Sealing Kit, for 96-well plates, includes optical compression pad, 100 Microseal B clear adhesive seals 223-9444 Optical Sealing Tape, package of 100 sheets 223-9442 96-Well PCR Plate Sealing Mats, 5Amplification Reagents and Plastics 34 Visit us on the Web at www.bio-rad.com.
Cy is a trademark of GE Healthcare group companies. Eppendorf and Mastercycler are trademarks of Eppendorf AG. EvaGreen is a trademark of Biotium, Inc. Bio-Rad Laboratories, Inc. islicensed by Biotium, Inc. to sell reagents containing EvaGreen dye for use in real-time PCR, for research purposes only. FAM, ROX, StepOne, StepOnePlus, Veriti, and VIC are trademarks ofApplera Corporation. HRM and Rotor-Gene are trademarks of QIAGEN GmbH. LightCycler is a trademark of Roche Diagnostics GmbH. LightScanner is a trademark of Idaho Technology Inc. Mx,Mx3000P, Mx3005P, and Mx4000 are trademarks of Stratagene Corporation. SYBR is a trademark of Molecular Probes, Inc. Bio-Rad Laboratories, Inc. is licensed by Molecular Probes, Inc. to sellreagents containing SYBR Green I for use in real-time PCR, for research purposes only. Texas Red is a trademark of Invitrogen Corporation. ViiA is a trademark of Life Technologies Corporation.Notice regarding Bio-Rad thermal cyclers and real-time systems:Purchase of this instrument conveys a limited non-transferable immunity from suit for the purchaser’s own internal research and development and for use in human in vitro diagnostics and all otherapplied fields under U.S. Patent Number 5,475,610 (Claims 1, 44, 158, 160–163, and 167 only), or corresponding claims in its non-U.S. counterpart, owned by Applera Corporation. No right is conveyedexpressly, by implication, or by estoppel under any other patent claim, such as claims to apparatus, reagents, kits, or methods such as 5 nuclease methods. Further information on purchasing licensesmay be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.Bio-Rad’s real-time thermal cyclers are licensed real-time thermal cyclers under Applera’s U.S. Patent Number 6,814,934 B1 for use in research, human in vitro diagnostics, and all other fields exceptveterinary diagnostics.Bio-Rad’s thermal cyclers and real-time thermal cyclers are covered by one or more of the following U.S. patents or their foreign counterparts owned by Eppendorf AG: U.S. Patent Numbers 6,767,512and 7,074,367.Practice of the patented 5 Nuclease Process requires a license from Applied Biosystems. The purchase of these products includes an immunity from suit under patents specified in the product insertto use only the amount purchased for the purchaser’s own internal research when used with the separate purchase of Licensed Probe. No other patent rights are conveyed expressly, by implication, orby estoppel. Further information on purchasing licenses may be obtained from the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.Hard-Shell plates are covered by one or more of the following U.S. patents or their foreign counterparts owned by Eppendorf AG: U.S. Patent Numbers 7,347,977; 6,340,589; and 6,528,302. Bio-Rad Laboratories, Inc. Life Science Web site www.bio-rad.com USA 800 424 6723 Australia 61 2 9914 2800 Austria 01 877 89 01 Belgium 09 385 55 11 Brazil 55 31 3689 6600 Canada 905 364 3435 China 86 21 6169 8500 Czech Republic 420 241 430 532 Denmark 44 52 10 00 Finland 09 804 22 00 Group France 01 47 95 69 65 Germany 089 31 884 0 Greece 30 210 777 4396 Hong Kong 852 2789 3300 Hungary 36 1 459 6100 India 91 124 4029300 Israel 03 963 6050 Italy 39 02 216091 Japan 03 6361 7000 Korea 82 2 3473 4460 Malaysia 60 3 2117 5260 Mexico 52 555 488 7670 The Netherlands 0318 540666 New Zealand 64 9 415 2280 Norway 23 38 41 30 Poland 48 22 331 99 99 Portugal 351 21 472 7700 Russia 7 495 721 14 04 Singapore 65 6415 3170 South Africa 27 861 246 723 Spain 34 91 590 5200 Sweden 08 555 12700 Switzerland 061 717 95 55 Taiwan 886 2 2578 7189 Thailand 66 2 6518311 United Kingdom 020 8328 2000 Bulletin 6090 Rev B US/EG 11-1398 0911 Sig 0211