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:1. DNA DNA template plasmid5-20 ng 　　10x pfu DNA polymerase buffer5.0 µl 　　25uM oligo 10.5 µl 　　25uM oligo 20.5 µl 　　10mM dNTP1.0 µl 　　Pfu DNA polymerase (2.5 units)1.0 µ[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]

摘要:1. For double-stranded DNA templates:a. Denature template: 10ml DNA (5-10g or alkaline lysis mini prepDNA)8ml ddH2O　　2ml 2N NaOH　　-incubate 30' at 37℃.b. Dry-ice precipitate: +10ml 4M NH4OAc100ml EtOH[阅读全文]

摘要:一、质粒的提取及鉴定（一）质粒的提取及鉴定1、收获细菌（1）将2ml含相应抗生素的LB液体培养基加入到通气良好的15ml的试管中，接入一单菌落，于37℃剧烈振荡培养过夜。（2）将1.5ml培养物倒入1.5ml离心管中，用台式离心机于4℃以1[阅读全文]

摘要:Below we present the protocols we have used to isolate DNA from various tissues using Silica and Guanidinium Thiocyanate.These protocols are adapted from Boom et al. (1990),Höss & Pääbo[阅读全文]

摘要:Phenol (removes protein) 1.add equal volume of Phenol (= tris-saturated Phenol-Chloroform-Isoamyethanol) 2.vortex 3.spin 2 minutes at 12000 rpm 4℃4.transfer supernatant to a fresh tube (avoid aspirati[阅读全文]

摘要:I am seeking procedures and/or advice on how to isolate human genomic DNA from platoregistry sera. The sera is lymphocytes that have been frozen for 35-40 years but are not dessicated.Will the isolate[阅读全文]

摘要:Abstract: Single strand conformational polymorphism (SSCP)is the most widely used PCR-based methods for point mutation detection. The abnormal band found by SSCP analysis is normally verified using se[阅读全文]

摘要:Tissue collection, storage, microdissection, sectionin g: See separate protocol.Tissue handling : Note that all fresh tissue shoμld be handled as BioSafety Level 2 materials (wear gloves, lab coat,[阅读全文]

摘要:- make a 5µm section to do an evaluation of the % tumour cells- make 50X50µm sections for the DNA isolation- put sections in a falcon tube containing 10 ml of 1XSE- add 100µg/ml prot[阅读全文]

摘要:Does anyone have expereince of extracting DNA from urine? I am having problems with this. I have previously extracted the DNA but then it degraded very quickly and since then I havent even been able t[阅读全文]

摘要:Caltech Genome Research Laboratory,California Institute of Technology http://www.tree.caltech.edu/protocols/agfp.html Restriction digests consist of:15.75 ml ddH2O 1 µl 10 X buffer B (Boehringer[阅读全文]

摘要:I usually use Linear polyacrylamide as a carrier for DNA precipitation by EtOH. I have read about using glycogen as a carrier, I would like to know if anyone knows a method avoiding not to use acrylam[阅读全文]