Haplotype-Contained PCR Products Analysis by Sequencing with Selective Restriction of Primer Extension
Joint Authors
Source
Issue
Vol. 2017, Issue 2017 (31 Dec. 2017), pp.1-9, 9 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2017-12-10
Country of Publication
Egypt
No. of Pages
9
Main Subjects
Abstract EN
We develop a strategy for haplotype analysis of PCR products that contained two adjacent heterozygous loci using sequencing with specific primers, allele-specific primers, and ddNTP-blocked primers.
To validate its feasibility, two sets of PCR products, including two adjacent heterozygous SNPs, UGT1A1⁎6 (rs4148323) and UGT1A1⁎28 (rs8175347), and two adjacent heterozygous SNPs, K1637K (rs11176013) and S1647T (rs11564148), were analyzed.
Haplotypes of PCR products, including UGT1A1⁎6 and UGT1A1⁎28, were successfully analyzed by Sanger sequencing with allele-specific primers.
Also, haplotypes of PCR products, including K1637K and S1647T, could not be determined by Sanger sequencing with allele-specific primers but were successfully analyzed by pyrosequencing with ddNTP-blocked primers.
As a result, this method is able to effectively haplotype two adjacent heterozygous PCR products.
It is simple, fast, and irrespective of short read length of pyrosequencing.
Overall, we fully hope it will provide a new promising technology to identify haplotypes of conventional PCR products in clinical samples.
American Psychological Association (APA)
Wang, Liu& Xiao, Pengfeng. 2017. Haplotype-Contained PCR Products Analysis by Sequencing with Selective Restriction of Primer Extension. BioMed Research International،Vol. 2017, no. 2017, pp.1-9.
https://search.emarefa.net/detail/BIM-1133880
Modern Language Association (MLA)
Wang, Liu& Xiao, Pengfeng. Haplotype-Contained PCR Products Analysis by Sequencing with Selective Restriction of Primer Extension. BioMed Research International No. 2017 (2017), pp.1-9.
https://search.emarefa.net/detail/BIM-1133880
American Medical Association (AMA)
Wang, Liu& Xiao, Pengfeng. Haplotype-Contained PCR Products Analysis by Sequencing with Selective Restriction of Primer Extension. BioMed Research International. 2017. Vol. 2017, no. 2017, pp.1-9.
https://search.emarefa.net/detail/BIM-1133880
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1133880