Journal of Visualized Experiments

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Pyrosequencing: A Simple Method for Accurate Genotyping
Cristi King, Tiffany Scott-Horton
Department of Internal Medicine, Washington University of St. Louis

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0:00 Title0

0:40 Introduction40

2:19 Preparing the PCR Product for Pyrosequencing139

6:42 Pyrosequencing402

6:45 Entering Assay Details Using Pyrosequencing 96A1.1 Software405

8:36 Run Sample Plate516

11:16 Examining Pyrosequencing Results676

12:18 Conclusion738

Pharmacogenetic research benefits first-hand from the abundance of information provided by the completion of the Human Genome Project. With such a tremendous amount of data available comes an explosion of genotyping methods. Pyrosequencing(R) is one of the most thorough yet simple methods to date used to analyze polymorphisms. It also has the ability to identify tri-allelic, indels, short-repeat polymorphisms, along with determining allele percentages for methylation or pooled sample assessment. In addition, there is a standardized control sequence that provides internal quality control. This method has led to rapid and efficient single-nucleotide polymorphism evaluation including many clinically relevant polymorphisms. The technique and methodology of Pyrosequencing is explained.

The complete text protocol for this experimental approach is available in Springer Protocols.

King C, Scott-Horton T (2008). Pyrosequencing: A Simple Method for Accurate Genotyping. JoVE. 11. http://www.jove.com/index/details.stp?id=630, doi: 10.3791/630
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08/18/2008 11:45:51 PM
joss responded with a statement of type: Neutral

thank's for u'r share. this is usefull.

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03/28/2010 7:16:18 AM
qqsvery responded with a statement of type: Agree
hello!your video is great ~ I can not get the complete text protocol for this experimental in Springer Protocols, could you please send it to my e-mail :qqsvery@hotmail.com. I am a postgraduate from JiangXi Agriculture University in China .thanks for your help !
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01/08/2008

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doi: 10.3791/630 

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Journal of Visualized Experiments (JoVE) is an online research journal employing visualization to increase reproducibility and transparency in biological sciences.

 

ISSN 1940-087X

 

 

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