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SCHOTT Jenaer Glas GmbH
Technical Support
Phosphate Buffer based Spotting Solutions
Slide E Slide A Slide
A+
Slide H Slide...
SCHOTT Jenaer Glas GmbH
Technical Support
Low Evaporation Spotting Buffers
Slide E Slide A Slide
A+
Slide H Slide
AL
25% t...
SCHOTT Jenaer Glas GmbH
Technical Support
DMSO containing Spotting Buffers on
Slide E
Spot Size
SMP3
Spot Size aQu
K2801
2...
SCHOTT Jenaer Glas GmbH
Technical Support
Adjust Spot Size for DMSO containing
Spotting Solutions
data shown on previous ...
SCHOTT Jenaer Glas GmbH
Technical Support
Slide E Slide A Slide
A+
Slide H Slide
AL
Comments
3x SSC     
3x SSC + 1.5...
SCHOTT Jenaer Glas GmbH
Technical Support
Slide E Slide A Slide
A+
Slide H Slide
AL
Comments
3x SSC     
3x SSC + 1.5...
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Compatibility of different Spotting / Print buffers with Nexterion Slide coatings

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  • last update 3.12.2004
  • Spotter: QArray Mini (Genetix)
    Print Head for aQu pins: Genetix HPLF head
    Print Head for SMPs pins: Telechem print head
    Probe: 70mer (Operon, f.c. 10 µM)
    Target: Dye 3(5) labeled cDNA
  • Transcript of "Compatibility of different Spotting / Print buffers with Nexterion Slide coatings"

    1. 1. SCHOTT Jenaer Glas GmbH Technical Support Phosphate Buffer based Spotting Solutions Slide E Slide A Slide A+ Slide H Slide AL Comments Nexterion Spot (phosphate buffer based)      add detergent to increase spot size1 50 to 300 mM Phosphate Buffer, pH 8.0 to 9.0      50 mM phosphate buffer = CodeLink Buffer 300 mM Phosphate Buffer + 0.005% Tween20 + 0.001% sarcosyl      Spot H (50) 150 mM Phosphate Buffer + 0.001% Tween20      Spot H (30) 1 detergents: SDS, sarcosyl, Tween, Triton, Cetyl-trimethylammoniumbromid, final concentrations needs to be determined empirically, usual f.c. range between 0.001% and 0.05%
    2. 2. SCHOTT Jenaer Glas GmbH Technical Support Low Evaporation Spotting Buffers Slide E Slide A Slide A+ Slide H Slide AL 25% to 50% ethylene glycol in phosphate buffer  ? - ? - 25% to 50% ethylene glycol in H2O - ? - ?  25% to 50% DMSO in H2O -   - - 25% to 50% DMSO in phosphate buffer - ? ? 2 ? 3x SSC + 1.5 M betaine      2 used by a customer
    3. 3. SCHOTT Jenaer Glas GmbH Technical Support DMSO containing Spotting Buffers on Slide E Spot Size SMP3 Spot Size aQu K2801 25% DMSO, 1.5x SSC, 0.005% SDS* 90 µm 160 µm 30% DMSO, 1.5x SSC 70 µm 90 µm 50% DMSO, 1.5x SSC 70 µm - (inhomogeneous spots) 50% DMSO, 1.5x SSC, 0.005% SDS* 60-70 µm 110 µm 50(30) % DMSO, 2x or 2.5x SSC 100 to 130 µm * higher concentrations of SDS may lead to precipitation, other detergents also work: Triton X100, cetyl- trimethylammonium bromide, both with a f.c. of 0.025%) Problem: with DMSO as spotting solution for Slide E are small spot sizes (as low as 50 µm) Solution: add SSC or SSC plus detergent, do not add phosphate buffer or Next Spot
    4. 4. SCHOTT Jenaer Glas GmbH Technical Support Adjust Spot Size for DMSO containing Spotting Solutions data shown on previous slide are based on a final concentration of 10 µM (70mer, Operon) if a customer has Oligos already dissolved in DMSO, usually the final concentrations of oligos is 20 µM and higher to adjust spot size, SSC and SDS may be added: Example: customer has 50 µl of 20 µM oligo dissolved in 50% DMSO add: 50 µl of 50% DMSO containing 3x SSC and 0.01% SDS final concentration of components 10 µM oligo (sufficient for Slide E) 50% DMSO 1.5x SSC 0.005% SDS
    5. 5. SCHOTT Jenaer Glas GmbH Technical Support Slide E Slide A Slide A+ Slide H Slide AL Comments 3x SSC      3x SSC + 1.5 M betaine      low evaporation 50% DMSO in 0.5x SSC* 3 ? ? ? ? low evaporation SSC based Spotting Buffers 3 used by a customer, most probably will work also on Slide A, A+ * for other combinations of DMSO and SSC see also slide “DMSO containing spotting buffers on Slide E”
    6. 6. SCHOTT Jenaer Glas GmbH Technical Support Slide E Slide A Slide A+ Slide H Slide AL Comments 3x SSC      3x SSC + 1.5 M betaine      low evaporation 50% DMSO in 0.5x SSC* 3 ? ? ? ? low evaporation SSC based Spotting Buffers 3 used by a customer, most probably will work also on Slide A, A+ * for other combinations of DMSO and SSC see also slide “DMSO containing spotting buffers on Slide E”
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