Institute of Laboratory Medicine and Pathobiochemistry, Charité University Medicine Berlin, Augustenburger Platz 1, 13353, Berlin, Germany.
Biotechnol Lett. 2010 Mar;32(3):421-2. doi: 10.1007/s10529-009-0157-4. Epub 2009 Nov 7.
Commercially available plasmid preparation kits are used in many laboratories to isolate plasmid DNA for subsequent dideoxy sequencing. We tested kits from seven different manufacturers for the purity of obtained plasmid preparations and compared the quality of sequencing results. The amount of RNA carry-over differed significantly (80-323 ng) and the usage of two kits led to protein remainders. Sequence read lengths (0-812 bases) and percentages of reading errors (1.6-100%) were also significantly different. We found the amount of protein contaminants to correlate with the percentage of reading errors (r = 0.63, P = 0.02).
商业上可获得的质粒制备试剂盒在许多实验室中用于分离质粒 DNA，以便随后进行双脱氧测序。我们测试了来自 7 个不同制造商的试剂盒，以评估获得的质粒制备物的纯度，并比较了测序结果的质量。RNA 残留量差异显著（80-323ng），有两个试剂盒会导致残留蛋白。序列读取长度（0-812 个碱基）和读取错误百分比（1.6-100%）也有显著差异。我们发现蛋白质污染物的含量与读取错误百分比相关（r=0.63，P=0.02）。
