Advances and Clinical Practice in Pyrosequencing (Springer by Guohua Zhou, Qinxin Song

By Guohua Zhou, Qinxin Song

The goal of this ebook is to enhance pyrosequencing protocols in addition to instrumentation for higher medical use by way of describing advancements and novel functions of pyrosequencing technology.  Divided into 5 elements, the book’s thirty chapters discover advances in pyrosequencing template training, pyrosequencing know-how suggestions, multiplex pyrosequencing in keeping with barcodes, the miniaturization of pyrosequencing apparatus, in addition to numerous functions. As a part of the Springer Protocols application, chapters include the type of element and useful implementation suggestion to assure winning ends up in the lab.

Comprehensive and thorough, Advances and scientific perform in Pyrosequencing serves as a helpful reference for researchers who're engaged in customized drugs, disorder keep an eye on, and DNA analysis in different different fields.

Show description

Read or Download Advances and Clinical Practice in Pyrosequencing (Springer Protocols Handbooks) PDF

Similar anatomy books

Title Cognitive Foundations of Grammar

The most functionality of language is to show which means. The query of why language is dependent how it is, Heine right here argues, has as a result to be responded firstly just about this functionality. Linguistic factors when it comes to different exponents of language constitution, e. g. of syntax, tend to spotlight peripheral or epi-phenomenal instead of principal features of language constitution.

A.D.A.M. Interactive Anatomy Online Student Lab Activity Guide

The ADAM Interactive Anatomy on-line: scholar Lab job advisor is geared to assist deliver much more which means and alertness to the cloth you’re studying on your Anatomy & body structure path.  No subject what allied well-being self-discipline you’re getting ready for, this advisor can help convey the cloth to lifestyles, make the content material extra significant to the genuine global, and position you at the route to mastery of human anatomy and body structure.

Nucleic Acid Aptamers: Selection, Characterization, and Application (Methods in Molecular Biology)

This quantity offers protocol references masking fresh advancements within the aptamer box. in the final decade, aptamers became increasingly more renowned, and their subtle biophysical homes including their skill to be simply transformed and, hence, tailored to varied regimens makes them a truly promising category of compounds.

Additional resources for Advances and Clinical Practice in Pyrosequencing (Springer Protocols Handbooks)

Sample text

For the advantages of accuracy, flexibility, and parallel processing [2–5], pyrosequencing has been widely used for DNA resequencing [6–8], genotyping [9, 10], DNA methylation [11], and gene expression analysis [12]. In conventional pyrosequencing protocol, a biotin-modified PCR primer and streptavidin-coated beads are required to prepare ssDNA template prior to pyrosequencing. 1007/978-1-4939-3308-2_4, © Springer Science+Business Media New York 2016 31 32 Qinxin Song et al. prepare ssDNA are expensive [13] and multiple steps that are needed to prepare the template increase the risk of DNA contamination in the air.

2. PCR mixture denatured at 94 °C for 15 min and followed by 35 cycles (94 °C for 40 s; 55 °C for 40 s; 72 °C for 1 min). After the cycle reaction, the product was incubated at 72 °C for 10 min and held at 4 °C before use. 2 Treatment of dsDNA with Nicking Endonucleases 1. PCR products were purified by the commercially available PCR purification kit or enzymatically treated by apyrase coupled with ATP sulfurylase (see Note 1). 2. BstNBI in a 1× reaction buffer (see Note 2). 5–1 h with 5–10 U of nicking endonuclease (see Note 3).

Most importantly, the length of amplicons should be short enough to ensure that Tm difference between the amplicon and the excess primer is less than 13 °C. Usually, an SNP loci of interest is fixed, so a difficulty may occur in designing qualified LATE-PCR primers at GC-rich or AT-rich regions. The main problem of low amplification efficiency of LATE-PCR is due to the competition of excess primer with the amplicon strand. As amplicon strand is increasing in the concentration along with the progress of PCR, while the excess primer is decreasing in the concentration, the yield of ssDNA should not be high if the amplicon is longer [3–6].

Download PDF sample

Rated 4.06 of 5 – based on 40 votes