摘要:点击浏览该文件[阅读全文]

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: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:This is a standard large scale prep.for plasmid DNA which gives a yield of 0.5-1.0mg.I have made some minor changes to the MHB protocol.Solutions Solution I,II,III from protocol D.1.Tris/EDTA pH 7.5 ([阅读全文]

摘要:1. set up the following reaction: CIP RxnH2 O 7.8 ml 10x cip rxn buffer 2.0 ml DNA(e.g; 3 kb vector; 0.2 mg/ml; 2 mg total) 10.0 ml (1 u/ml) CIP 0.2 ml total 20.0 ml [阅读全文]

摘要:This method is after Marchuk,D.,et al.,1991,Nucl.Acids Res.19(5),pp1154. You will need:10 x Taq buffer (Promega)Taq Polymerase (Promega)Phenol/chloroform mix100mM dTTPTE bufferAbsolute ethanol70% etha[阅读全文]

摘要:1) The ligation mixture contains the following:vector (~100 ng)　　insert (equimolar or 2 or 3 X molar concentration of vector)　　water added to 18 µl2) Heat the mixture at 45 ℃ for 5 min. to melt [阅读全文]

摘要:Preparation of Transformation Competent DNA Dilute an overnight bacterial culture 1:20 in L broth. (For E. coli strain JA221, at A )Using a serological pipet, resuspend the bacterial pellet gently in [阅读全文]

摘要:Procedure1. Grind 2 to 5 g of frozen leaves to a very fine powder with a liquid nitrogen-cooled mortar and pestle.2. Add 25 ml of CTAB Buffer and transfer to a 50 ml tube.3. Incubate at 65℃ for 20 min[阅读全文]

摘要:Caution: EtBr is a powerful mutagen and is moderately toxic. Wear gloves when working. When EtBr conc. is >0.5 mg/ml: 1.Add sufficient water to reduce the concentration of EtBr to <0.5 mg/ml. 2.Add 1 [阅读全文]

摘要:Hello group!I want to insert a small 30 bp sequence into a vector of mine using ds-oligo. I've created the oligos so that once annealed, they have the correct overhangs of the restriction sites.As I'm[阅读全文]

摘要:Hey folks,I need Help!! Please!I have to put a 3.5 kB fragment into a 6.5 kB vector.The vector has a Km resistance gene.I use invitrogen chemically competent cells.This is what I do:I PCR amplify the [阅读全文]

摘要:(adapted from Bruce A. Roe, Department of Chemistry and Biochemistry, The University of Oklahoma, Norman, Oklahoma 73019 broe@ou.edu)Typically, 2.5 - 3 volumes of an ethanol/acetate solution is added [阅读全文]

摘要:原理： 转化(Transformation )：将外源DNA 分子引入受体细菌，使之获得新的遗传性状。 受体细菌一般是限制修饰系统缺陷的变异株，不含限制性内切酶和甲基化酶的突变体(R-,M-)，可以容忍外源DNA 分子进入体内并稳定地遗传给[阅读全文]

摘要:CTAB TECHNIQUE / Method / Schedule / Protocol (JPB) FOR DNA ISOLATION / DNA EXTRACTION FROM PLANT LEAF / LEAVES SAMPLES(see also if both DNA and RNA are needed)Reagents neededCTAB buffer2% CTAB20gm CT[阅读全文]

摘要:Preparation of C. elegans Genomic DNAProtocol by Andy Fires LabModified my Min-Ho Lee and Sudhir NayakThis protocol is based on the original Fire lab protocol for isolating genomic DNA from C. elegans[阅读全文]

摘要:CpG islands can be variable in length. Anyone know by what percentage range they can typically vary? I can't find much of any herlp on Google.CheersChris -Chris Harris---------------------------------[阅读全文]

摘要:CaCl2 感受态细胞的制备的实验步骤1．前夜接种受体菌（ DH5α或 DH10B），挑取单菌落于LB培养基中37℃摇床培养过夜（约16小时）； 2．取 1ml过夜培养物转接于100ml LB培养基中，在37℃摇床上剧烈振荡培养约2.5-3小时（250-3[阅读全文]

摘要:Dr. William H. Heidcamp, Biology Department, Gustavus Adolphus College http://homepages.gac.edu/~cellab/chpts/chpt14/ex14-6.html Materials•DNA•CsCl•0.3 N NaOH•0.2 M Tris-HCl buffer[阅读全文]

摘要:In the hood:96 well dish with bacteriatitertechmicrotubesglass pipetteRemove colonies from each well using the titertech and place them into the coverPipette up and down to thoroughly mix the colonies[阅读全文]