会议专题

PREPARATION OF SINGLE-STRANDED DNA FOR PYROSEQUENCING BY6 USING LINEAR-AFTER-THE-EXPONENTIAL (LATE)-PCR

The preparation of single-stranded (ssDNA) DNA template is important for pyrosequencing. Nowdays a new asymmetric PCR amplification method (named imLATE-PCR) is described in this paper; imLATE-PCR can also prepare ssDNA of high quality by improving the amplification efficiency of asymmetric PCR. The method guarantees the amplification efficiency and specialization via by altering the the reaction annealing temperature and using PCR enhancer during the exponential phase. During the phase of linear amplification, the imLATE-PCR has higher ssDNA amplification efficiency and can amplify longer PCR products than LATE-PCR for its higher melting temperature (Tm) of the excess primer (PX). The ssDNA templates which prepared from the products of imLATE-PCR, the sequences of SNP1, SNP2 and HA gene of Avian influenza virus, were pyrosequenced. The results indicate that imLATE-PCR can prepare high quality ssDNA templates for pyrosequencing as well as have excellent effect in pyrogram, cost, operation and is easier in automation.

Yang Huiyong Zhou Guohua

Huadong Research Institute for Medicine and Biotechnics, Nanjing, China Huadong Research Institute for Medicine and Biotechnics, Nanjing, China Medical School, Nanjing Univ

国际会议

The 6th International Forum on Post-genome Technologies(6IFPT)(第六届国际后基因组生命科学技术学术论坛)

北京

英文

403-407

2009-09-17(万方平台首次上网日期,不代表论文的发表时间)