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An efficient cost‐effective protocol for automated fluorescent‐DNA sequencing
Author(s) -
Beg Obaid U.,
Holt Robert G.
Publication year - 1997
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1111/j.1470-8744.1997.tb00443.x
Subject(s) - dna sequencing , dna , sequencing by ligation , sequencing by hybridization , computational biology , dna nanoball sequencing , sequence (biology) , protocol (science) , chemistry , biology , base sequence , dna sequencer , genetics , genomic library , medicine , alternative medicine , pathology
A simple and cost‐effective method is described to sequence double‐stranded DNA (< 100 ng) using a very small amount of fluorescent sequencing mix (one‐tenth of manufacturer‐recommended amount). The modification has significant advantages over conventional protocols and can be used to sequence, using a single sequencing kit, at least ten times the number of samples. This modified method also removes the post‐reaction cleaning protocol and can be used for direct loading of samples on gels after completion of sequencing reactions. The accuracy of DNA sequencing data from analysis of 700 to 750 nucleotides is greater than 98.5′.