Goelet P, Karn J
Gene. 1984 Sep;29(3):331-42. doi: 10.1016/0378-1119(84)90062-3.
DNA complementary to tobacco mosaic virus (TMV) RNA (cDNA) was prepared by priming reverse transcription with synthetic oligonucleotides. The cDNAs terminated prematurely at many specific sites and no transcripts longer than about 2000 nucleotides were obtained. However, the entire 6395 nucleotide long TMV RNA could be copied into cDNA by specific priming with a series of 13 to 17 residue long oligonucleotides or by non-specific priming with short, 4 to 7 residue, oligonucleotides. A number of different priming methods were used to convert the cDNA into double-stranded DNA (dsDNA). The double-stranded cDNA was recovered by shotgun cloning into M13 and analysed by sequencing. The frequency at which cDNA clones were recovered has been used to compare various cDNA cloning strategies.
通过用合成寡核苷酸引发逆转录制备与烟草花叶病毒（TMV）RNA互补的DNA（cDNA）。cDNA在许多特定位点过早终止，未获得长度超过约2000个核苷酸的转录本。然而，通过用一系列13至17个残基长的寡核苷酸进行特异性引发或用短的4至7个残基的寡核苷酸进行非特异性引发，可将全长6395个核苷酸的TMV RNA复制到cDNA中。使用了多种不同的引发方法将cDNA转化为双链DNA（dsDNA）。通过鸟枪法克隆到M13中回收双链cDNA并进行测序分析。回收cDNA克隆的频率已用于比较各种cDNA克隆策略。
