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Titolo:
Directional cloning of native PCR products with preformed sticky ends (Autosticky PCR)
Autore:
Gal, J; Schnell, R; Szekeres, S; Kalman, M;
Indirizzi:
Bay Zoltan Fdn Appl Res, Inst Biotechnol, H-6701 Szeged, Hungary Bay Zoltan Fdn Appl Res Szeged Hungary H-6701 ol, H-6701 Szeged, Hungary Hungarian Acad Sci, Biol Res Ctr, Inst Genet, H-6701 Szeged, Hungary Hungarian Acad Sci Szeged Hungary H-6701 t Genet, H-6701 Szeged, Hungary
Titolo Testata:
MOLECULAR AND GENERAL GENETICS
fascicolo: 6, volume: 260, anno: 1999,
pagine: 569 - 573
SICI:
0026-8925(199901)260:6<569:DCONPP>2.0.ZU;2-O
Fonte:
ISI
Lingua:
ENG
Soggetto:
POLYMERASE CHAIN-REACTION; TRANSLESION DNA-SYNTHESIS; SINGLE ABASIC SITE; ENZYMATIC AMPLIFICATION; DEOXYRIBONUCLEIC-ACID; EFFICIENT CLONING; ESCHERICHIA-COLI; APURINIC SITES; GENERATED DNA; A-RULE;
Keywords:
PCR cloning; sticky ends; abasic primer;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
40
Recensione:
Indirizzi per estratti:
Indirizzo: Gal, J Bay Zoltan Fdn Appl Res, Inst Biotechnol, POB 2337, H-6701 Szeged, Hungary Bay Zoltan Fdn Appl Res POB 2337 Szeged Hungary H-6701 ed, Hungary
Citazione:
J. Gal et al., "Directional cloning of native PCR products with preformed sticky ends (Autosticky PCR)", MOL G GENET, 260(6), 1999, pp. 569-573

Abstract

A novel method for the directional cloning of native PCR products was developed. A basic sites in DNA templates make DNA polymerases stall at the site during synthesis of the complementary strand. Since the 5' ends of PCR product strands contain built-in amplification primers, abasic sites within the primers result in the formation of 5' single-stranded overhangs at the ends of the PCR product, enabling its direct ligation to a suitably cleaved cloning vector without any further modification. This "autosticky PCR" (AS-PCR) overcomes the problems caused by end sensitivity of restriction enzymes, or internal restriction sites within the amplified sequences, and enables the generation of essentially any desired 5' overhang.

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Documento generato il 05/12/20 alle ore 02:01:45