Automated DNA Sequencing Requiring No DNA Template Purification

A critical component in automated fluorescent DNA sequencing is good quality of DNA template. In an effort to reduce the dependence of sequencing success on DNA template quality, the effect of heat-soaked PCR on automated sequencing reactions has been examined. We have found that the heat-soaked PCR...

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Main Authors: Qianfa Chen, Catherine Neville, Alex MacKenzie, Robert G. Korneluk
Format: Article
Language:English
Published: Taylor & Francis Group 1996-09-01
Series:BioTechniques
Online Access:https://www.future-science.com/doi/10.2144/96213st02
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author Qianfa Chen
Catherine Neville
Alex MacKenzie
Robert G. Korneluk
author_facet Qianfa Chen
Catherine Neville
Alex MacKenzie
Robert G. Korneluk
author_sort Qianfa Chen
collection DOAJ
description A critical component in automated fluorescent DNA sequencing is good quality of DNA template. In an effort to reduce the dependence of sequencing success on DNA template quality, the effect of heat-soaked PCR on automated sequencing reactions has been examined. We have found that the heat-soaked PCR protocol considerably improves the overall quality of sequence data and significantly reduces the dependence on the quality of DNA templates. The improvement is corroborated by our ability to obtain over 500 bp of readable sequence per reaction using DNA from E. coli lysates as template obviating DNA purification.
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publishDate 1996-09-01
publisher Taylor & Francis Group
record_format Article
series BioTechniques
spelling doaj-art-ff4aeb45aafe4adeb0f54a5975eb319a2025-08-20T02:26:06ZengTaylor & Francis GroupBioTechniques0736-62051940-98181996-09-0121345345710.2144/96213st02Automated DNA Sequencing Requiring No DNA Template PurificationQianfa Chen0Catherine Neville1Alex MacKenzie2Robert G. Korneluk31Children's Hospital of Eastern, Ontario, Ottawa, ON, Canada1Children's Hospital of Eastern, Ontario, Ottawa, ON, Canada1Children's Hospital of Eastern, Ontario, Ottawa, ON, Canada1Children's Hospital of Eastern, Ontario, Ottawa, ON, CanadaA critical component in automated fluorescent DNA sequencing is good quality of DNA template. In an effort to reduce the dependence of sequencing success on DNA template quality, the effect of heat-soaked PCR on automated sequencing reactions has been examined. We have found that the heat-soaked PCR protocol considerably improves the overall quality of sequence data and significantly reduces the dependence on the quality of DNA templates. The improvement is corroborated by our ability to obtain over 500 bp of readable sequence per reaction using DNA from E. coli lysates as template obviating DNA purification.https://www.future-science.com/doi/10.2144/96213st02
spellingShingle Qianfa Chen
Catherine Neville
Alex MacKenzie
Robert G. Korneluk
Automated DNA Sequencing Requiring No DNA Template Purification
BioTechniques
title Automated DNA Sequencing Requiring No DNA Template Purification
title_full Automated DNA Sequencing Requiring No DNA Template Purification
title_fullStr Automated DNA Sequencing Requiring No DNA Template Purification
title_full_unstemmed Automated DNA Sequencing Requiring No DNA Template Purification
title_short Automated DNA Sequencing Requiring No DNA Template Purification
title_sort automated dna sequencing requiring no dna template purification
url https://www.future-science.com/doi/10.2144/96213st02
work_keys_str_mv AT qianfachen automateddnasequencingrequiringnodnatemplatepurification
AT catherineneville automateddnasequencingrequiringnodnatemplatepurification
AT alexmackenzie automateddnasequencingrequiringnodnatemplatepurification
AT robertgkorneluk automateddnasequencingrequiringnodnatemplatepurification