摘要:1. Pick single colony and inoculate 5 ml of LB broth containing 200 g/l ampicillin or 1mg/5ml. Optional: Use a 15ml conical tube with a loosened cap and a piece of tape to hold it in place. Shake at 2[阅读全文]

摘要: 点击浏览该文件 [阅读全文]

摘要:reagents: DNA for labeling (concentration c > 150 ng/µl) modified nucleotides: Biotin-16-dUTP, Digoxigenin-11-dUTP, conc. 1nmol/µl (Boehringer Mannheim) dNTPs (regular nucleotides): dATP, [阅读全文]

摘要:DNA methylation is an epigenetic event that affects cell function by altering gene expression and refers to the covalent addition of a methyl group, catalyzed by DNA methyltransferase (DNMT), to the 5[阅读全文]

摘要:点击浏览该文件[阅读全文]

摘要:Sample Preparation Cheek cells are obtained by rinsing the mouth with 25mls of any commercial mouth wash solution available for about 30sec the first thing in the morning. It is important not to brush[阅读全文]

摘要:一、 DNA 酶切反应 1、将清洁干燥并经灭菌的eppendorf管(最好0.5ml)编号，用微量移液枪分别加入DNA 1μg和相应的限制性内切酶反应10×缓冲液2μl，再加入重蒸水使总体积为19μl，将管内溶液混匀后加入1μl酶[阅读全文]

摘要:在分子生物学研究中，DNA 的序列分析是进一步研究和改造目的基因的基础。目前用于测序的技术主要有Sanger等（1977）发明的双脱氧链末端终止法和Maxam和 Gilbert（1977）发明的化学降解法。这二种方法在原理上差异很大，但都[阅读全文]

摘要:Hahn Lab，The Fred Hutchinson Cancer Research Center and Howard Hughes Medical Institutehttp://www.fhcrc.org/science/labs/hahn/methods/mol_bio_meth/Big%20Dye%20Protocol.pdf [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.Prepare or obtain Buffered phenol,pH 8.0.Add 2 crystals of 8-Hydroxyquinoline to prevent oxidation.This also identifies the organic phase as yellow-colored.2.Combine DNA sample with an equal volume [阅读全文]

摘要:ReagentsChloroformEDTA,0.5 MEthanol, absoluteIsoamyl alcoholSigma,Cat.I-3643PhenolPhosphate Buffered Saline(PBS),1XProteinase KEM Science,Gibbstown,WV Cat.24568-2(100 mg)RNase ABoehringer Mannheim,Cat[阅读全文]

摘要:This is a scaled up version of the Baron protocol which has been modified to achieve purity comparable to CsCl preps. I have not sorted out strain variations yet but HB101 works well.1、SolutionsSoluti[阅读全文]

摘要:Phenol (removes protein)1、add equal volume of Phenol (= tris-saturated Phenol-Chloroform-Isoamyethanol)2、vortex3、spin 2 minutes at 12000 rpm 4℃4、transfer supernatant to a fresh tube (avoid aspiration [阅读全文]

摘要:Equipment and reagents1.Phenol2. TE buffer,pH 8.0 (10 mM Tris-HCl,pH 8.0;1 mM EDTA,pH 8.0)3. 24:1 (v/v) chloroform-isoamyl alcohol4. 3 M potassium acetate, pH 5.5,prepared by adding glacial acetic aci[阅读全文]

摘要:1. Separate DNA fragments in an agarose gel cast with 0.5 mg/ml Ethidium bromide. Locate bands with a hand-held long-wave UV lamp.2. Slice the gel with a razor blade above and below the bands of inter[阅读全文]

摘要:1) Take one medium sized leaf or half a large leaf (5 to 20 cm^2), weigh and freeze in liquid nitrogen.2) Grind the tissue in a bleached and baked pestle and mortar with liquid nitrogen.3) Transfer th[阅读全文]

摘要:DNA priming reaction x ul DNA2 ul Reaction buffer, minus DTT1 ul primer (20 ng)10 ul totalHeat 90-100C 2 min.Cool room temp. 30 min.Enzyme mix4 ul radioactive nucleotide (12 l)2 ul reaction buffer( 6 [阅读全文]

摘要:The sequencing reactions described below work perfectly well if you are short of cash to buy sequencing kits.It is based on the Dideoxy sequencing method of Sanger et al.,1977.However,due to the numbe[阅读全文]

摘要:Buffers and gel solutions Long Ranger: we started using this in early 1995. Great stuff; the best thing is that the gels are not sticky after drying, even without removal of the urea. Long Ranger Gel [阅读全文]