This document discusses three methods for detecting nucleic acids:
1. UV spectroscopy can be used to estimate the concentration and purity of DNA and RNA samples by measuring absorbance at 260nm and 280nm. Ratios below 1.8 indicate contamination.
2. Ethidium bromide staining allows nucleic acids to be visualized under UV light after gel electrophoresis, as the dye intercalates within DNA and RNA.
3. Fluorometric quantification uses dyes like Hoechst 33258 and DAPI that bind preferentially to DNA or RNA and fluoresce at different wavelengths, allowing for quantification.
Isolation and Purification of Chromosomal DNA,Plasmid DNA,Bacteriophage DNA used in Recombinant DNA Technology or Biotechnology to produce Recombinant DNA or Desired DNA
Nucleic Acid Quantification Methods - DNA / RNA Quantificationajithnandanam
Nucleic acids are quantified to check the concentration and purity of DNA/RNA present in the solution mixture.it is important to know the concentration and purity of the nucleic acid for the use in further applications like PCR, restriction digestion etc. Spectrophotometric analysis is the most commonly used method of quantifying DNA, agarose gel electrophoresis can also be used to analyse the DNA sample for purity.
Isolation and Purification of Chromosomal DNA,Plasmid DNA,Bacteriophage DNA used in Recombinant DNA Technology or Biotechnology to produce Recombinant DNA or Desired DNA
Nucleic Acid Quantification Methods - DNA / RNA Quantificationajithnandanam
Nucleic acids are quantified to check the concentration and purity of DNA/RNA present in the solution mixture.it is important to know the concentration and purity of the nucleic acid for the use in further applications like PCR, restriction digestion etc. Spectrophotometric analysis is the most commonly used method of quantifying DNA, agarose gel electrophoresis can also be used to analyse the DNA sample for purity.
A DNA library is a collection of cloned restriction fragments of the DNA of an organism.
Two kinds of libraries will be discussed: genomic libraries and complementary DNA (cDNA) libraries.
Genomic libraries ideally contain a copy of every DNA nucleotide sequence in the genome.
In contrast, cDNA libraries contain those DNA sequences that appear as mRNA molecules, and these differ from one cell type to another.
MBB 501 PLANT BIOTECHNOLOGY
INFORMATION ABOUT DIFFERENT DNA MODIFYING ENZYMES
WHAT IS AN ENZYME?
Alkaline Phosphatase
Polynucleotide kinase
Terminal deoxyneucleotidyl transferase
Nucleases
Exonuclease
Bal31 Exonuclease III
Endonuclease
S1 endonulease
Deoxyribonuclease 1 (Dnase 1)
RNase A
RNase H
Restriction Endonuclease
PvuI
PvuII
Different types of endonuclease enzymes
The recognition sequences for some of the most frequently used restriction endonucleases.
Categorization of enzymes
Isoschizomers
Neoschizomers
Isocaudomers
This presentation contains information about restriction enzymes, its nomenclature, restriction digestion, and its application. This also contains information about the chemicals used in restriction and also explains the general procedure of restriction digestion of DNA
A DNA library is a collection of cloned restriction fragments of the DNA of an organism.
Two kinds of libraries will be discussed: genomic libraries and complementary DNA (cDNA) libraries.
Genomic libraries ideally contain a copy of every DNA nucleotide sequence in the genome.
In contrast, cDNA libraries contain those DNA sequences that appear as mRNA molecules, and these differ from one cell type to another.
MBB 501 PLANT BIOTECHNOLOGY
INFORMATION ABOUT DIFFERENT DNA MODIFYING ENZYMES
WHAT IS AN ENZYME?
Alkaline Phosphatase
Polynucleotide kinase
Terminal deoxyneucleotidyl transferase
Nucleases
Exonuclease
Bal31 Exonuclease III
Endonuclease
S1 endonulease
Deoxyribonuclease 1 (Dnase 1)
RNase A
RNase H
Restriction Endonuclease
PvuI
PvuII
Different types of endonuclease enzymes
The recognition sequences for some of the most frequently used restriction endonucleases.
Categorization of enzymes
Isoschizomers
Neoschizomers
Isocaudomers
This presentation contains information about restriction enzymes, its nomenclature, restriction digestion, and its application. This also contains information about the chemicals used in restriction and also explains the general procedure of restriction digestion of DNA
This is technique used widely for protein separation from a mixture and is very easy and less costly method. Slides cover all essential points about EMSA and it is quite interesting to know that how it detect and separate different proteins and their mobility shift assay.
In this presentation you will get a deep insight on the most important step of DNA fingerprinting that is the Quantitation of DNA.
You will understand what is DNA quantitation and also about the different techniques of DNA quantitation.
Lab 23 DNA Extraction and PurificationIsolation and purific.docxDIPESH30
Lab 2/3: DNA Extraction and Purification
Isolation and purification of nucleic acids is the most fundamental procedure in molecular biology. There are three basic steps involved:
1. Lyse (break open) the cells (and nuclei in eukaryotes) to release the DNA
2. Remove contaminants (proteins, lipids, carbohydrates, salts)
3. Preserve the integrity of the DNA (prevent degradation and shearing)
Step 1 can be accomplished in a number of ways, such as mechanical disruption (grinding, mincing), protein denaturation (detergents), and protein degradation (via proteases). These can be used singly or in combination depending on the type of biological sample you are starting with. Grinding the samples provides more surface area for the denaturants/proteases to interact with the cellular proteins, thus speeding up the denaturation process. We used liquid nitrogen (N2) and protein degradation (Proteinase K) in lab 2. Various salts are included in a cell lysis solution to stabilize the DNA by providing positive ions which insert between the negatively charged phosphates in the DNA backbone (creating a “salt bridge”). Buffers (such as Tris) also help to preserve DNA integrity by maintaining a neutral pH.
Once the cells have been lysed, contaminating proteins, lipids, etc. must be separated from the DNA. A widely used and efficient way to remove proteins from nucleic acids solutions is to extract with a 1:1 mixture of phenol and chloroform (CHCl3). Phenol and CHCl3 are both hydrophobic organic solvents that unfold proteins. When mixed with an aqueous DNA/protein solution and then centrifuged, the denatured proteins are selectively partitioned into the denser organic phase, while the DNA (plus RNA and salt) remains in the aqueous phase. This procedure takes advantage of the fact that deproteinization is more efficient when two different organic solvents are used instead of one. Additionally, chloroform removes any lingering traces of phenol from the nucleic acid preparation (which would interfere with later applications). Since the aqueous phase contains RNA and salt in addition to the DNA, phenol:CHCl3 extraction is followed by ethanol (EtOH) precipitation. DNA (a polar molecule) is soluble in water (also polar) because the water molecules intercalate into the phosphate backbone of the DNA and thus maintain it in a soluble state, but DNA is insoluble in 95% EtOH (nonpolar). Water molecules have a higher affinity for the EtOH than the DNA, so when you add EtOH and salt [10 M ammonium acetate (NH4Ac); pH 5.2], Na+ ions replace water in the DNA backbone, essentially removing the water molecules, and the DNA is forced out of solution (precipitates). After precipitating with 95% EtOH, the DNA is “washed” in 70% EtOH to remove the salt. Since 70% EtOH contains 30% water, the salt, having a greater affinity for the water than the DNA, remains in the EtOH, and the DNA is forced out.
The final step in the purification process is to preserve the DNA in a sta ...
this section helps students how to quanify the isolated DNA by spectrophotometer. specially life life science fields such as biotechnology, biology, and medical laboratory
liposomes and nanoparticles drug delivery systemShreyaBhatt23
this presentation includes the intro duction to targeted drug delivery systems using nanoparticulate systems like liposomes, nanoparticles, mechanism of action, types, preparation, advantages, applications
experiences and outcomes of the internship done at VI solutions the presentation contains the brief introduction to LabVIEW and tasks fullfilled at workspace, conclusion,references
Nanotechnology in cancer and its synthesisShreyaBhatt23
basic introduction to nanotechnology and the types of nanomaterials used in medical purpose. sysnthesis of nanomaterials by physical , chemical, biosynthesis, green synthesis of nanomaterials
Introduction to nanoparticles and bionanomaterialsShreyaBhatt23
what is a nanoparticle, why small is good,nanoscale effect, how to make nanostructures,top down and bottom up approachs,
methods of making nanomaterials,chemical methods od making nanomaterial,bionanomaterials,
HIGH PERFORMANCE LIQUID CHROMATOGRAPHY(HPLC)ShreyaBhatt23
HIGH PERFORMANCE LIQUID CHROMATOGRAPHY workflow, types of hplc, normal phase HPLC, reverse phase HPLC,types of column, advantages pf HPLC over other liquid chromatography, parameters of HPLC. SOURCE: ARPAN YOUTUBE
biocompatibility of biopolymers and their sterilisation techniques.ShreyaBhatt23
what is biopolymers, types of biopolymers, classification of biopolymers, natural biopolymers, sterilization techniques of polymers like dry heating, autoclaving, radiation , chemical agents
Enzymes of biological importance LDH and 5' NTShreyaBhatt23
clinical importance of the enzymes lactate dehydrogenase and 5' neucleotidase . various assays used to treat disorders, basic knowledge about the enzymes used in clinical chemistry aspects and their saient features
taking example of aprotein sequence luciferan 4-monoxygenase i have shown the uniprotkb/swiss prot format database for bioinformatics laboratory that is sequence retrieval
Homo sapiens (human pepsin) NCBI GENBANKShreyaBhatt23
GenBank format and FASTA format as homo sapiens pepsin as an example bioinformatics practical 1st experiment ; sequence retrival from nucleotide sequence from NCBI
Overview of the fundamental roles in Hydropower generation and the components involved in wider Electrical Engineering.
This paper presents the design and construction of hydroelectric dams from the hydrologist’s survey of the valley before construction, all aspects and involved disciplines, fluid dynamics, structural engineering, generation and mains frequency regulation to the very transmission of power through the network in the United Kingdom.
Author: Robbie Edward Sayers
Collaborators and co editors: Charlie Sims and Connor Healey.
(C) 2024 Robbie E. Sayers
Explore the innovative world of trenchless pipe repair with our comprehensive guide, "The Benefits and Techniques of Trenchless Pipe Repair." This document delves into the modern methods of repairing underground pipes without the need for extensive excavation, highlighting the numerous advantages and the latest techniques used in the industry.
Learn about the cost savings, reduced environmental impact, and minimal disruption associated with trenchless technology. Discover detailed explanations of popular techniques such as pipe bursting, cured-in-place pipe (CIPP) lining, and directional drilling. Understand how these methods can be applied to various types of infrastructure, from residential plumbing to large-scale municipal systems.
Ideal for homeowners, contractors, engineers, and anyone interested in modern plumbing solutions, this guide provides valuable insights into why trenchless pipe repair is becoming the preferred choice for pipe rehabilitation. Stay informed about the latest advancements and best practices in the field.
Automobile Management System Project Report.pdfKamal Acharya
The proposed project is developed to manage the automobile in the automobile dealer company. The main module in this project is login, automobile management, customer management, sales, complaints and reports. The first module is the login. The automobile showroom owner should login to the project for usage. The username and password are verified and if it is correct, next form opens. If the username and password are not correct, it shows the error message.
When a customer search for a automobile, if the automobile is available, they will be taken to a page that shows the details of the automobile including automobile name, automobile ID, quantity, price etc. “Automobile Management System” is useful for maintaining automobiles, customers effectively and hence helps for establishing good relation between customer and automobile organization. It contains various customized modules for effectively maintaining automobiles and stock information accurately and safely.
When the automobile is sold to the customer, stock will be reduced automatically. When a new purchase is made, stock will be increased automatically. While selecting automobiles for sale, the proposed software will automatically check for total number of available stock of that particular item, if the total stock of that particular item is less than 5, software will notify the user to purchase the particular item.
Also when the user tries to sale items which are not in stock, the system will prompt the user that the stock is not enough. Customers of this system can search for a automobile; can purchase a automobile easily by selecting fast. On the other hand the stock of automobiles can be maintained perfectly by the automobile shop manager overcoming the drawbacks of existing system.
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Welcome to WIPAC Monthly the magazine brought to you by the LinkedIn Group Water Industry Process Automation & Control.
In this month's edition, along with this month's industry news to celebrate the 13 years since the group was created we have articles including
A case study of the used of Advanced Process Control at the Wastewater Treatment works at Lleida in Spain
A look back on an article on smart wastewater networks in order to see how the industry has measured up in the interim around the adoption of Digital Transformation in the Water Industry.
Final project report on grocery store management system..pdfKamal Acharya
In today’s fast-changing business environment, it’s extremely important to be able to respond to client needs in the most effective and timely manner. If your customers wish to see your business online and have instant access to your products or services.
Online Grocery Store is an e-commerce website, which retails various grocery products. This project allows viewing various products available enables registered users to purchase desired products instantly using Paytm, UPI payment processor (Instant Pay) and also can place order by using Cash on Delivery (Pay Later) option. This project provides an easy access to Administrators and Managers to view orders placed using Pay Later and Instant Pay options.
In order to develop an e-commerce website, a number of Technologies must be studied and understood. These include multi-tiered architecture, server and client-side scripting techniques, implementation technologies, programming language (such as PHP, HTML, CSS, JavaScript) and MySQL relational databases. This is a project with the objective to develop a basic website where a consumer is provided with a shopping cart website and also to know about the technologies used to develop such a website.
This document will discuss each of the underlying technologies to create and implement an e- commerce website.
Saudi Arabia stands as a titan in the global energy landscape, renowned for its abundant oil and gas resources. It's the largest exporter of petroleum and holds some of the world's most significant reserves. Let's delve into the top 10 oil and gas projects shaping Saudi Arabia's energy future in 2024.
Water scarcity is the lack of fresh water resources to meet the standard water demand. There are two type of water scarcity. One is physical. The other is economic water scarcity.
Forklift Classes Overview by Intella PartsIntella Parts
Discover the different forklift classes and their specific applications. Learn how to choose the right forklift for your needs to ensure safety, efficiency, and compliance in your operations.
For more technical information, visit our website https://intellaparts.com
COLLEGE BUS MANAGEMENT SYSTEM PROJECT REPORT.pdfKamal Acharya
The College Bus Management system is completely developed by Visual Basic .NET Version. The application is connect with most secured database language MS SQL Server. The application is develop by using best combination of front-end and back-end languages. The application is totally design like flat user interface. This flat user interface is more attractive user interface in 2017. The application is gives more important to the system functionality. The application is to manage the student’s details, driver’s details, bus details, bus route details, bus fees details and more. The application has only one unit for admin. The admin can manage the entire application. The admin can login into the application by using username and password of the admin. The application is develop for big and small colleges. It is more user friendly for non-computer person. Even they can easily learn how to manage the application within hours. The application is more secure by the admin. The system will give an effective output for the VB.Net and SQL Server given as input to the system. The compiled java program given as input to the system, after scanning the program will generate different reports. The application generates the report for users. The admin can view and download the report of the data. The application deliver the excel format reports. Because, excel formatted reports is very easy to understand the income and expense of the college bus. This application is mainly develop for windows operating system users. In 2017, 73% of people enterprises are using windows operating system. So the application will easily install for all the windows operating system users. The application-developed size is very low. The application consumes very low space in disk. Therefore, the user can allocate very minimum local disk space for this application.
1. METHODS OF NUCLEIC ACID
DETECTION
NAME : KALPANA LAMANI
USN : 2BA18BT005
DEPARTMENT OF BIOTECHNOLOGY
2. 1. UV SPECTROSCOPY
• This property of UV absorbance can be used quickly to estimate
the concentration and purity of DNA and RNA in analytical
sample.
• The relationship between concentration of DNA,RNA,protein
and absorptivity are as below :
• The purity of a solution of nucleic acid is determined by
measuring the absorbance of the solution at two wavelengths,
usually 260nm and 280nm, and calculating the ratio of
3. • Value of this ratio is 2.0,1.8 and 0.6 for pure RNA,DNA and protein
respectively. A ratio of less than 1.8 signifies that the sample is
contaminated with protein or phenol and the preparation is not proper.
Fig : UV detection of nucleic acid
Fig : Nucleic acid melting
curve
showing hyperchromicity
4. 2. ETHIDIUM BROMIDE STAINING
It is commonly used as fluorescent dye for nucleic acid staining. It binds
as well as intercalates with nucleic acid and gives orange fluorescence
under UV radiation from 500nm-590nm. Usually EtBr may be added in
warm agarose gel before solidification.When DNA or RNA samples are run
in agarose gel electrophoresis EtBr molecules will bind with nucleic acids
and helps in detection under UV light.
5. 3. FLUOROMETRIC QUANTIFICATION
An assay using Hoechst 33258 dye is specific for DNA because it
is less sensitive to detect RNA. Hoechst 33258 is non intercalating
reagent and binds to the minor groove of the DNA with preference
for AT sequences. This dye is positively charged and thus form
electrostatic interaction with the negative potential of stretch of
AT base pairs. Upon binding to the minor groove of the double
helix DNA,the fluorescence characteristics of Hoechst 33258
change dramatically, showing a large increase in emission at
458nm
DAPI (4’,6-diamidino-2-phenylindole) has the similar
characteristics to H33258 is also appropriate for DNA or RNA
quantitaion,although it is not as commonly used as Hoechst
33258 DAPI is excited with a peak at 344nm. Emission is detected