A DNA cycle sequencing reaction that minimizes compressions on automated fluorescent sequencers | Zendy

Hisashi Yamakawa | Zendy

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A DNA cycle sequencing reaction that minimizes compressions on automated fluorescent sequencers

Author(s) -

Hisashi Yamakawa

Publication year - 1997

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/25.6.1311

Subject(s) - biology , dna sequencing , dna , dna sequencer , dna nanoball sequencing , base pair , massive parallel sequencing , microbiology and biotechnology , computational biology , genetics , base sequence , genomic library

We recently demonstrated that most band compressions (>70%) on DNA sequencing gels result from the presence of a single sequence motif, 5'-YGN1-2AR-3', where Y and R indicate base-pairing pyrimidine and purine residues, respectively. This finding raised the possibility that the use of 7-deaza-dATP instead of dATP in chain termination sequencing reactions would resolve most of the band compressions. Thus, we examined the effects of 7-deaza-dATP on DNA sequencing using thermostable DNA polymerases. The results indicate that the replacement of dATP with 7-deaza-dATP in conventional cycle sequencing reactions can successfully eliminate most band compressions without sacrificing sequencing performance.

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