Substrate nucleotide-determined non-templated addition of adenine by Taq DNA polymerase: Implications for PCR-based genotyping and cloning

被引：134

作者：

Magnuson, VL ^{[1
]}

Ally, DS ^{[1
]}

Nylund, SJ ^{[1
]}

Karanjawala, ZE ^{[1
]}

Rayman, JB ^{[1
]}

Knapp, JI ^{[1
]}

Lowe, AL ^{[1
]}

Ghosh, S ^{[1
]}

Collins, FS ^{[1
]}

机构：

[1] PERKIN ELMER CORP,APPL BIOSYST DIV,FOSTER CITY,CA

来源：

BIOTECHNIQUES | 1996年 / 21卷 / 04期

关键词：

D O I：

10.2144/96214rr03

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The Applied Biosystems PRISM(TM) florescence-based genotyping system as well as the Invitrogen TA Cloning(R) vector system are influenced by the tendency of Taq DNA polymerase to add an adenine nucleotide to the 3' end of PCR products after extension. Incomplete addition of adenine to a majority of PCR product strands creates problems in allele-calling during genotyping and potentially diminishes the cloning efficiency of such products. Experiments reported here show that certain terminal nucleotides can either inhibit or enhance adenine addition by Taq and that PCR primer design can be used to modulate this activity. The methods we propose can substantially improve allele-calling for problematic microsatellite markers when using GENOTYPER(TM) software.

引用

页码：700 / 709

页数：10

共 9 条

[1] NOVEL NON-TEMPLATED NUCLEOTIDE ADDITION-REACTIONS CATALYZED BY PROCARYOTIC AND EUKARYOTIC DNA-POLYMERASES [J].