Efficient reamplification of differential display products by transient ligation and thermal asymmetric PCR

被引：7

作者：

Bonnet, S ^{[1
]}

Prévot, G ^{[1
]}

Bourgouin, C ^{[1
]}

机构：

[1] Inst Pasteur, Unite Ecol Syst Vectoriels, F-75015 Paris, France

来源：

NUCLEIC ACIDS RESEARCH | 1998年 / 26卷 / 04期

关键词：

D O I：

10.1093/nar/26.4.1130

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A new method for specific reamplification of DDRT-PCR products is presented. After transient ligation of the primary DDRT-PCR fragments into a T-vector, the cDNAs of interest were reamplified by hemi-nested PCR and thermally asymmetric cycles. In contrast to the originally described protocol, this method of reamplification is specific, sensitive, reproducibly gives a high yield of DNA and allows direct sequencing of the reamplified product without purification or cloning.

引用

页码：1130 / 1131

页数：2

共 17 条

[1]

CALLARD D, 1994, BIOTECHNIQUES, V16, P1096

[2]

CHOMCZYNSKI P, 1993, BIOTECHNIQUES, V15, P532

[3]

Chomczynski P, 1993, BIOTECHNIQUES, V15, P536

[4] Identification and characterization of differentially expressed cDNAs of the vector mosquito, Anopheles gambiae [J].

Dimopoulos, G ;

Richman, A ;

dellaTorre, A ;

Kafatos, FC ;

Louis, C .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (23) :13066-13071

[5] Rational primer design greatly improves differential display PCR (DD-PCR) [J].

Graf, D ;

Fisher, AG ;

Merkenschlager, M .

NUCLEIC ACIDS RESEARCH, 1997, 25 (11) :2239-2240

[6] ISOPROPANOL PRECIPITATION REMOVES PCR INHIBITORS FROM ANCIENT BONE EXTRACTS [J].

HANNI, C ;

BROUSSEAU, T ;

LAUDET, V ;

STEHELIN, D .

NUCLEIC ACIDS RESEARCH, 1995, 23 (05) :881-882

[7] DIFFERENTIAL DISPLAY OF EUKARYOTIC MESSENGER-RNA BY MEANS OF THE POLYMERASE CHAIN-REACTION [J].

LIANG, P ;

PARDEE, AB .

SCIENCE, 1992, 257 (5072) :967-971

[8] RECENT ADVANCES IN DIFFERENTIAL DISPLAY [J].

LIANG, P ;

PARDEE, AB .

CURRENT OPINION IN IMMUNOLOGY, 1995, 7 (02) :274-280

[9]

LIU C, 1995, PLANT PHYSIOL, V109, P1126

[10] THERMAL ASYMMETRIC INTERLACED PCR - AUTOMATABLE AMPLIFICATION AND SEQUENCING OF INSERT END FRAGMENTS FROM P1 AND YAC CLONES FOR CHROMOSOME WALKING [J].

LIU, YG ;

WHITTIER, RF .

GENOMICS, 1995, 25 (03) :674-681

← 1 2 →