摘要:目的：在体外连接组装而成的重组DNA分子只能转入合适的受体细胞，才能大量地进行复制、增殖和表达。通过实验学会重组DNA转化的最基本的操作以及如何提高转化效率的基本思路。原理：带有外源DNA片段的重组体分子在体外构建[阅读全文:]
摘要:Preparation of plasmid DNA : a modified mini alkaline-lysis/PEG precipitation procedure. ABI; 1995MaterialsGTE buffer (50mM glucose, 25mM Tris-HCl (pH8.0), 10mM EDTA (pH 8.0)) (200µl/tube) 0.2N [阅读全文:]
摘要:Simple and Efficient Method (SEM) to Make Competent CellsPreparation of Frozen Competent of DH5α1) 250 ml TB solution 10mM Pipes or 10 mM Hepes, 0.65g 15 mM CaCl2, 0.55g 250 mM KCl, 4.66g *55 mM[阅读全文:]
摘要:Hello,I have been trying over the last few months to introduce a single point mutation in a looped portion of an RNA, using the dut-ung method of mutagenesis.So far, I have had little success.I have a[阅读全文:]
摘要:I tried to clean a restriction product >10kb before ligation by a gel extraction method (Qiagen gel extraction kit). But the concentration was too low to do a ligation. I used to clean restriction pro[阅读全文:]
摘要:I have tried to purify my PCR products by using 3 different approaches:1) QIAQuick PCR purification kit: but when I run the gel to see its efficacy, there was simply nothing!! it cleaned everything in[阅读全文:]
摘要:The Preuss Lab,The Division of Biological Sciences,The University of Chicago http://preuss.bsd.uchicago.edu/protocols/topo.html [阅读全文:]
摘要:LiAc/SS-DNA/PEG TRANSFORMATIONTRAFO Efficiency: up to 22 million transformanst/ µg of plasmid DNAHIGH EFFICIENCY TRANSFORMATIONPlease cite:Agatep, R., R.D. Kirkpatrick, D.L. Parchaliuk, R.A. Woo[阅读全文:]