scholarly journals Rapid sequencing of unpurified PCR products by thermal asymmetric PCR cycle sequencing using unlabeled sequencing primers

1993 ◽  
Vol 21 (14) ◽  
pp. 3333-3334 ◽  
Author(s):  
Yao-Guang Liu ◽  
Norihiro Mitsukawa ◽  
Robert F. Whittier
Keyword(s):  
Asymmetric Pcr ◽  
Cycle Sequencing ◽  
Pcr Products

scholarly journals Hepatitis B Transmission: Is Vertical Transmission the Major Route in Intermediate Endemic Areas? A Proof-of-Concept Study Based on Mother–Child Genotypes

2021 ◽  
Author(s):  
Ujjal Poddar ◽  
Mercilena Benjamin ◽  
Rakesh Aggarwal ◽  
Aditya Narayan Sarangi ◽  
Amrita Mathias ◽  
...  
Keyword(s):  
Hepatitis B ◽  
Region Of Interest ◽  
Cycle Sequencing ◽  
Child Pair ◽  
Mother And Child ◽  
Pcr Products ◽  
Major Route ◽  
Genetic Sequences ◽  
Hepatitis B Transmission ◽  
Genotype A

The route of hepatitis B transmission is believed to be horizontal in India, though pediatric studies showed mother as source in the majority of chronic HBV (CHB) cases. We aimed at establishing the fact that mother–child transmission is the main route of acquisition by documenting genotypically identical viruses in mother–child pairs. Blood samples of consecutive children (≤18 years) with CHB and high DNA (>10,000 IU/mL) and their positive mother were collected from January 2013 to December 2015. Polymerase chain reaction (PCR) products of HBV-DNA were amplified and sequenced by using BigDye Terminator Cycle Sequencing Kit v3.1 and aligned with previously described sequences in the region of interest for genotypes A to G by using BioEdit software. Phylogenetic tree was generated using p-distance algorithm in MEGA software version 6. Genotyping of 59 (33 children and 26 mothers) subjects include genotype A in 24 (40.7%) and genotype D in 35 (59.3%). Both mother–child pair genotyping was possible in 25. The median age of 25 children (20 males) was 9 (interquartile range, IQR: 4–11). The distribution of genotypes among mother–child pairs was similar. The concordance between children and their mothers was 24 of 25 (96%). Evolutionary analyses showed significant similarities between mother and child sequences for both genotype A and D, suggesting thereby the same virus. In conclusion, mother–baby transmission seems to be the major route of acquisition of HBV in children in India and near-complete homology in genetic sequences between mother–child pairs is definite proof for that. However, a larger epidemiological study is required to substantiate our findings.

Electrochemical Detection of Asymmetric PCR Products by Labeling with Osmium Tetroxide

2009 ◽  
pp. NA-NA ◽  
Author(s):  
Maren Mix ◽  
Thomas Reske ◽  
Heiko Duwensee ◽  
Gerd-Uwe Flechsig
Keyword(s):  
Electrochemical Detection ◽  
Osmium Tetroxide ◽  
Asymmetric Pcr ◽  
Pcr Products

A universal method for directional cloning of PCR products based on asymmetric PCR

2009 ◽  
Vol 52 (1) ◽  
pp. 41 ◽  
Author(s):  
Bao-Li Wang ◽  
Yan-Li Jiao ◽  
Xiao-Xia Li ◽  
Fang Zheng ◽  
Hui Liang ◽  
...  
Keyword(s):  
Universal Method ◽  
Asymmetric Pcr ◽  
Directional Cloning ◽  
Pcr Products

Identification at strain level of Rhizoctonia solani AG4 isolates by direct sequence of asymmetric PCR products of the ITS regions

1996 ◽  
Vol 29 (2) ◽  
pp. 174-181 ◽  
Author(s):  
M. Boysen ◽  
Marisé Borja ◽  
Catalina del Moral ◽  
Oscar Salazar ◽  
V. Rubio
Keyword(s):  
Rhizoctonia Solani ◽  
Strain Level ◽  
Direct Sequence ◽  
Asymmetric Pcr ◽  
Its Regions ◽  
Pcr Products

scholarly journals Comparison of Sequencing by Hybridization and Cycle Sequencing for Genotyping of Human Immunodeficiency Virus Type 1 Reverse Transcriptase

2000 ◽  
Vol 38 (7) ◽  
pp. 2715-2721 ◽  
Author(s):  
George J. Hanna ◽  
Victoria A. Johnson ◽  
Daniel R. Kuritzkes ◽  
Douglas D. Richman ◽  
Javier Martinez-Picado ◽  
...  
Keyword(s):  
Amino Acids ◽  
Human Immunodeficiency Virus ◽  
Drug Resistance ◽  
Reverse Transcriptase ◽  
Population Based ◽  
Cycle Sequencing ◽  
Immunodeficiency Virus ◽  
Pcr Products ◽  
Hybridization Sequencing

The performances of two methods of nucleotide sequencing were compared for the detection of drug resistance mutations in human immunodeficiency virus type 1 reverse transcriptase (RT) in viruses isolated from highly RT inhibitor-experienced individuals. Of 11,677 amino acids deduced from population PCR products by both cycle sequencing and sequencing by hybridization to high-density arrays of oligonucleotide probes, 97.4% were concordant by both methods, 0.8% were discordant, and 1.7% had an ambiguous determination by at least one method. A higher rate of discordance (3.9%) was observed among RT inhibitor resistance-associated codons. In 45% of the isolates, RT codon 67 was deduced as the wild-type Asp by hybridization sequencing but as the zidovudine resistance-associated Asn by cycle sequencing. In other resistance-associated codon discordances, cycle sequencing also more commonly called a known resistance-associated amino acid than hybridization sequencing did. The nucleotide sequence in the vicinity of several codons with discordant calls influenced population-based hybridization sequencing. For isolates evaluated by additional sequencing of molecular clones of PCR products by both methods, the discordance between methods was less frequent (0.4% of all 5,994 amino acids and 0 of 494 drug resistance-associated codons). At positions which were discordant or ambiguous in the population sequences, the results of sequencing of clones by both methods were usually in agreement with the population cycle sequencing result. In summary, most RT codons were highly concordant by both methods of population-based sequencing, with discordances due in large part to genetic mixtures within or adjacent to discordant codons.

scholarly journals Optimized conditions for cycle sequencing of PCR products.

1994 ◽  
Vol 3 (6) ◽  
pp. 359-360 ◽  
Author(s):  
J H Horton ◽  
M D Hagen ◽  
M S Ko
Keyword(s):  
Cycle Sequencing ◽  
Pcr Products ◽  
Optimized Conditions
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