Five-color-based high-information-content fingerprinting of bacterial artificial chromosome clones using type IIS restriction endonucleases.

Abstract:

:We have developed a high-information-content fingerprinting (HICF) system for bacterial artificial chromosome (BAC) clones using a Type IIS restriction endonuclease, HgaI, paired with a Type II restriction endonuclease, RsaI. In the method described, unknown five-base overhangs generated with HgaI are partially or fully sequenced by modified fluorescent dideoxy terminators. Using an in-lane size standard labeled with a fifth dye, fragments are characterized by both the size and the sequence of its terminal one to five bases. The enhanced information content associated with this approach significantly increases the accuracy and efficiency of detecting shared fragments among BAC clones. We have compared data obtained from this method to predicted HICF patterns of 10 fully sequenced BACs. We have further applied HICF to 555 BAC clones to assemble contigs spanning 16p11.2 to 16p13.1 of human chromosome 16.

journal_name

Genomics

journal_title

Genomics

authors

Ding Y,Johnson MD,Chen WQ,Wong D,Chen YJ,Benson SC,Lam JY,Kim YM,Shizuya H

doi

10.1006/geno.2001.6547

subject

Has Abstract

pub_date

2001-06-01 00:00:00

pages

142-54

issue

2

eissn

0888-7543

issn

1089-8646

pii

S0888-7543(01)96547-4

journal_volume

74

pub_type

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