A reliable amplification technique for the characterization of genomic DNA sequences flanking insertion sequences.
A reliable amplification technique for the characterization of genomic DNA sequences flanking insertion sequences.
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一种可靠的扩增技术，用于表征插入序列侧翼的基因组 DNA 序列。
DOI：
10.1111/j.1574-6968.1998.tb12803.x
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发表时间：
1998
影响因子：
2.1
通讯作者：
Guilhot,C
中科院分区：
文献类型：
--
作者：
Prod'hom,G;Lagier,B;Pelicic,V;Hance,AJ;Gicquel,B;Guilhot,C
一种简单有效的连接介导的PCR（LMPCR）被描述用于扩增与已知序列相邻的DNA。该方法使用一种对已知序列特异的引物和第二种对连接至限制性基因组DNA的合成接头特异的引物。Perkin-Elmer AmpliTaq Gold聚合酶用于最小化非特异性引物退火和扩增。该LMPCR方法成功地应用于分离转座子诱变产生的分枝杆菌突变体中存在的移动的元件侧翼的DNA序列。
A simple and efficient ligation-mediated PCR (LMPCR) is described for amplifying DNA adjacent to known sequences. The method uses one primer specific for the known sequence and a second specific for a synthetic linker ligated to restricted genomic DNA. Perkin-Elmer AmpliTaq Gold polymerase is used to minimize non-specific primer annealing and amplification. This LMPCR method was successfully applied to isolate DNA sequences flanking mobile elements present in mycobacterial mutants generated by transposon mutagenesis.