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: | , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Taylor & Francis Group
1996-09-01
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| Series: | BioTechniques |
| Online Access: | https://www.future-science.com/doi/10.2144/96213st02 |
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| _version_ | 1850151935831703552 |
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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. |
| format | Article |
| id | doaj-art-ff4aeb45aafe4adeb0f54a5975eb319a |
| institution | OA Journals |
| issn | 0736-6205 1940-9818 |
| language | English |
| 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 |