摘要:This is a rapid method for chemical DNA sequencing which is commonly used as ladder for footprinting reactions or for sequencing of short DNA oligonucleotides.Reference: Bencini et al. (1984) Biotechn[阅读全文:]
摘要:Miniprep of BAC DNA1.Inoculate 4 - 8 tubes of 3ml/tube LB with 15μg/ml chloramphenicol with BAC clone and grow oveernight. Miniprep with AutoGen 740 according to the protocol described in our Web p[阅读全文:]
摘要:点击浏览该文件[阅读全文:]
摘要:METHOD:1.Isolate chromosomal DNA by using the CTAB protocols in current Pr M R.2.Make a CsCl preparation of pLAFr3 (or other appropriate cosmid).3.Cut pLAFr with BamHI phenol ext. + ETOH preparation.4[阅读全文:]
摘要:It is best to do a test ligation without insert, if the background is low, it would not be necessary to screen colonies for insert.1.colony PCR 1) Pick a colony from plate and suspend the colony in 10[阅读全文:]
摘要:The preferred method of de-phosphorylation uses the buffer system described by Pharmacia for most of their restriction endonucleases. You will need:10 x OPA+ Buffer (100mM Tris.acetate, pH 7.5, 100mM [阅读全文:]
摘要:单拷贝克隆产量低，高拷贝克隆稳定性差，一直让研究者在克隆 表达时难以选择。蛋白的表达就有诱导表达系统，可以实现人为地控制蛋白的表达时间和表达量——现在连质粒的拷贝数可以借助诱导的方法来人为控制了！[阅读全文:]
摘要: 一、DNA 的酶切与连接 （1）酶切反应：同质粒DNA 的鉴定，只不过是质粒DNA 换为载体DNA 。若大量酶切，则成比例增加。（2）加2倍体积的预冷无水乙醇和1/10体积的3mol/l NaAc混匀，-20℃2h以上。（3）15000rpm离心15min，弃上清。（4）[阅读全文:]
摘要:1、Oligo design（1）The 5' end (the homology arm) - choose 42 or more (we usually choose about 50) nts for the homology arms from the target DNA sequence simply according to where you want to insert the [阅读全文:]
摘要:一、In a microcentrifuge tube prepare a solution of linear DNA (25-50ng) in deionized water or TE buffer (10-35µl). 二、Add: （一）10X ligation buffer 5µl, （二）50% PEG 4000 solution (for blunt[阅读全文:]