Minimum GC-Rich Sequences for Overlap Extension PCR and Primer Annealing
Minimum GC-Rich Sequences for Overlap Extension PCR and Primer Annealing
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DOI：
10.1007/978-1-62703-764-8_12
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发表时间：
2014-01-01
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影响因子：
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通讯作者：
Akada, Rinji
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文献类型：
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作者：
Nakamura, Mikiko;Suzuki, Ayako;Akada, Rinji
PCR是从模板产生所需DNA片段的常用方法。用于PCR的寡核苷酸引物必须含有通常为20-30个核苷酸长并且与模板DNA的一部分相同的退火序列。然而，引物通常在其5'端含有额外的序列，其是限制性酶切位点、重组靶向序列或用于融合PCR的重叠序列。当这些额外的序列连接到它们的退火序列时，退火序列可以缩短。在这里，我们描述了通用的GC丰富的退火序列，可用于重叠延伸PCR和简单的框内添加所需的序列。
PCR is a common method to produce desired DNA fragments from templates. The oligonucleotide primers used for PCR must contain annealing sequences that are usually 20-30 nucleotides long and identical to a part of template DNA. However, primers often contain additional sequences at their 5' ends, which are restriction enzyme sites, recombination targeting sequences, or overlap sequences for fusion PCR. When these additional sequences are attached to their annealing sequences, the annealing sequences can be shortened. Here, we describe universal GC-rich annealing sequences useful for overlap extension PCR and simple in-frame addition of desired sequences.