Publish Your Research Online
Get Recognition - International Audience
Request for an Author Account | Login | Submit Article
|HOME||FAQ||TOP AUTHORS||FORUMS||PUBLISH ARTICLE|
Common Q&A for SequencingBY: Sherry Green | Category: Bioinformatics | Submitted: 2015-10-14 20:48:59
Article Summary: "This is an article about the common question for sequencing. I select them and I think they are useful for your doubt..."
With the development of all kinds of research, sequencing gradually plays a more important role in the lab.Here I list several common questions that will be needs in the process of sequencing.
Q: What are necessary for sequencing primers?
A: The specificity can be combined with template. The mismatch can not be more than for basic groups. It can not contain mix-bases and the length of bases should be above 17 and below 25. It should have high purity and not be dissolved by TE buffer.
Q: What kind of solution is better for DNA sample dissolution?
A: It's better to choose sterilized distilled water for dissolution. The sequencing reaction of DNA is also the polyreaction of Taq polymerase, which needs a most suitable condition of enzyme reaction. Someone may choose the TE buffer for the dissolution. Indeed, if we choose the buffer, the stability will be increased during the the period of preservation. However, it will lower the polymer properties after sequencing.
Q: What kind of DNA sample form is better?
A: We recommend thallus. It can remain the stability of the sample. If you really want to provide a DNA sample, pay more attention to the quantity and purity of sample, especially PCR products.
Q: If I choose the thallus, what form of thatllus is better?
A: The common forms include plate culture, stab culture, glycerol extender and fresh bacterium solution. It's better to choose stab culture or fresh bacterium. It's really inconvenient for the transportation of plate culture. The petri dish is fragile and the customer needs to send the sample again. The glycerol extender is more easily polluted than other methods, so it is also not a good option.
Q: How to preserve synthesized primers?
A: Undissolved primer is stable enough to preserve for at least one year with the temperature of -20?. And dissolved primers can be preserved after attenuation.
Q: Why my PCR primer can not be used for sequencing?
A: Not all kinds of PCR primers are suitable for sequencing. Here are inappropriate types:
Degenerate primer. Degenerate primer has several binding sites, which will influence the result of sequencing.
Random primer, such as the RAPD Primer. It can not fit perfectly with the template.
Overlong primer. The purity can not be guaranteed if the primer is too long. Besides, it more easily has several binding sites under a low reaction condition.
Primer with specific marks, which is the primer with fluorescence labeling. It has interference on the basic group of sequencing. Besides, primers with a great amount of marker genes are also not suitable for sequencing. It will have effects on the mobility of DNA fragments.
Impure primer. It will make noises of sequencing.
Q: What will be needed for PCR direct sequencing?
A: Specific amplification for PCR amplification and single brand is needed. And it must be purified by gel extraction. Besides, the purity of DNA is between 1.6 and 2.0; the concentration of DNA is 50ug/ml above.
Q: How many times can I do PCR reaction by 2OD primers?
A: Generally, about 400 times.
About Author / Additional Info:
Sherry Green, from CD Genomics, http://www.cd-genomics.com , a biotechnology company which provides sequencing, genotyping, microarray service for global researchers.
Comments on this article: (1 comments so far)
• Tank Bottom Sludge- Hazards and Treatment Methods
• Virtual Screening- a Promising Approach to Drug Discovery
• Deadly and Toxic Reptiles, Amphibians and Arachnids
• Progress of Protection of Plant Varieties and Farmers Rights Authority During 2006-07 to 2010-11
Latest Articles in "Bioinformatics" category:
• Career as Bioinformatician and Biostatistician
• Expander: A Tool of Bioinformatics
• Role of Bioinformatics in Drug Discovery
• Importance and Applications of Bioinformatics in Molecular Medicine
• Bioinformaticist vs. Bioinformatician - Definition, Differences and Career Outlook
• Bioinformatics Application in Nanotechnology
• How Bioinformatics Handles the Biological Data?
• Application of Bioinformatics in Medicine
• Prenatal Diagnosis via Bioinformatics Skills
• Applications of Bioinformatics in Agriculture
• Next Generation Sequencing Technologies: 454 Pyrosequencing
• GenScan: Bioinformatics Software For Structure Prediction and Analysis of Gene
• Pairwise Sequence Alignment For Sequence Similarity
• Applications of Bioinformatics in Biotechnology
• Introduction to Bioinformatics: Role of Mathematics and Technology
• Why and How of Normalization in Microarray Data Analysis
• Steps in Microarray Data Analysis - Part I
• Steps in Microarray Data Analysis - Part II
• Bilirubin Metabolism And its Role in Neonatal Jaundice
Important Disclaimer: All articles on this website are for general information only and is not a professional or experts advice. We do not own any responsibility for correctness or authenticity of the information presented in this article, or any loss or injury resulting from it. We do not endorse these articles, we are neither affiliated with the authors of these articles nor responsible for their content. Please see our disclaimer section for complete terms.
Copyright © 2010 biotecharticles.com - Do not copy articles from this website.
ARTICLE CATEGORIES :
| Disclaimer/Privacy/TOS | Submission Guidelines | Contact Us