摘要:1. Remove 0.5 cm of tail into polypropylene microfuge tube (do not mince).(The tubes must have tight-fitting caps, so that there are no leaks in steps 3 and 7 below.)2. Add 0.5 ml DNA digestion buffer[阅读全文]

摘要:Phenol-chloroform DNA extraction from sand flies1. Crash Individual sand fliesin 0.5 ml lysis buffer (50 mM NaCl , 10 mM EDTA, 50 mM Tris-HCl pH 8). 2.Incubate The sand fly homogenateswith 100 ng/ml R[阅读全文]

摘要:1. Remove gel slice contain DNA fragment and place in 10 volumes of:300 mM NaOAc, pH 7.0 300 ml 1 M NaOAC, pH 7.01 mM EDTA 2 ml 500 mM EDTA, pH 8.0698 ml ddH2O2. Incubate at 22 ℃ for 30 min. Transfer [阅读全文]

摘要:Diatomaceous Earth-based Midi-prep Note: This procedure is the method of choice for isolating double stranded plasmid-based templates for the Sequenase Dye-Labeled Terminator Sequencing Reactions. A. [阅读全文]

摘要:序列测定的技术和策略Sanger双脱氧链终止法Maxam－Gilbert DNA 化学降解法测序策略目前应用的两种快速序列测定技术是Sanger等（1977）提出的酶法及Maxam和Gilbert(1977)提出的化学降解法。虽然其原理大相径庭，但这两种方[阅读全文]

摘要: 点击浏览该文件 [阅读全文]

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:Molecularpourous membrane tubing is used for desalting, protein and DNA isolation / purification. It comes in several diameters and pore sizes (for molecular weight conversions for nucleic acids see g[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: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: 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]