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: 点击浏览该文件 [阅读全文:]
摘要:Restriction enzymes are expensive ($40 to $200 a vial): keep the enzymes at -20℃. Plan your digests to be small and convenient. The digest is composed of DNA sample (volume should not be more than abo[阅读全文:]
摘要:20cm Gel Preparation, Sample Separation, and Visualization: Glass Plate Preparation:[阅读全文:]
摘要:1)probe: same as used for gelshift), isolated by isotacelectrophoresis w/out EtOH ppt which can enature dsDNA 2)probe mix/rxn: volumes x # samples 1μl probe (0.1?0.5ng or 10?20kcpm) 0.15μl 20m[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: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:This protocol gives M13 quality sequence when used with the Quick DNA Plasmid Prep. protocol D.2.1、SolutionsDenaturation Solution 2 M NaOH 200 ml 10N NaOH 2 mM EDTA 4 ml 0.5 M EDTA pH 8.0 up to 1 m[阅读全文:]
摘要: 10X TBE, per literSequencing dye-----------------------------------108g Tris base80% formamide55g boric acid10mM NaOH40ml 0.5M EDTA1mM EDTA0.1% xylene cyanol10% APS0.1% bromophenol blue--------------[阅读全文:]