DEVELOPMENT OF AN AUTOMATED PROCEDURE FOR FLUORESCENT DNA SEQUENCING

被引：27

作者：

WILSON, RK ^{[1
]}

CHEN, C ^{[1
]}

AVDALOVIC, N ^{[1
]}

BURNS, J ^{[1
]}

HOOD, L ^{[1
]}

机构：

[1] BECKMAN INSTRUMENTS INC,SPINCO DIV,PALO ALTO,CA 94303

来源：

GENOMICS | 1990年 / 6卷 / 04期

基金：

美国国家卫生研究院; 美国国家科学基金会;

关键词：

D O I：

10.1016/0888-7543(90)90497-I

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

We describe here the development of a procedure for complete automation of the dideoxynucleotide DNA sequencing chemistry using fluorescent dye-labeled oligonucleotide primers. This procedure combines rapid preparation of template DNA using a modification of the polymerase chain reaction, automation of the DNA sequencing reactions using a robotic laboratory workstation, and subsequent analysis of the fluorescent-labeled reaction products on a commercial automated fluorescent sequencer. Using this procedure, we were able to produce sufficient quantities of template DNA directly from bacterial colonies or bacteriophage plaques, perform the DNA sequencing reactions on these templates, and load the reaction products on the fluorescent DNA sequencer in a single work day. This scheme for automation of the fluorescent DNA sequencing method allows the fluorescent sequencer to be run at its full capacity every day and eliminates much of the labor required to obtain a high level of data output. Currently, we are able to perform and analyze 16 fluorescent-labeled reactions every day, with an average output of over 7000 bp per sequencer run. © 1990.

引用

页码：626 / 634

页数：9

共 19 条

[1] AUTOMATED DNA SEQUENCING - ULTRASENSITIVE DETECTION OF FLUORESCENT BANDS DURING ELECTROPHORESIS
ANSORGE, W
SPROAT, B
STEGEMANN, J
SCHWAGER, C
ZENKE, M
[J]. NUCLEIC ACIDS RESEARCH, 1987, 15 (11) : 4593 - 4602
[2] AVDALOVIC N, 1988, ROBAN004 BECKM DIV P
[3] BANKIER A, 1988, ROBAN011 BECKM DIV P
[4] CONTINUOUS, ONLINE DNA SEQUENCING USING OLIGODEOXYNUCLEOTIDE PRIMERS WITH MULTIPLE FLUOROPHORES
BRUMBAUGH, JA
MIDDENDORF, LR
GRONE, DL
RUTH, JL
[J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (15) : 5610 - 5614
[5] CONNELL C, 1987, BIOTECHNIQUES, V5, P342
[6] GOCAYNE JD, 1988, FASEB J, V2, pA1125
[7] GENERATION OF SINGLE-STRANDED-DNA BY THE POLYMERASE CHAIN-REACTION AND ITS APPLICATION TO DIRECT SEQUENCING OF THE HLA-DQA LOCUS
GYLLENSTEN, UB
ERLICH, HA
[J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (20) : 7652 - 7656
[8] OPTIMIZATION OF PARAMETERS IN A DNA SEQUENATOR USING FLUORESCENCE DETECTION
KAMBARA, H
NISHIKAWA, T
KATAYAMA, Y
YAMAGUCHI, T
[J]. BIO-TECHNOLOGY, 1988, 6 (07): : 816 - 821
[9] A SIMPLE AND RAPID PREPARATION OF M13 SEQUENCING TEMPLATES FOR MANUAL AND AUTOMATED DIDEOXY SEQUENCING
KRISTENSEN, T
VOSS, H
ANSORGE, W
[J]. NUCLEIC ACIDS RESEARCH, 1987, 15 (14) : 5507 - 5516
[10] MESSING J, 1983, METHOD ENZYMOL, V101, P20

← 1 2 →