摘要:Materials: 1.TENS solution: 10 mM Tris (pH to 7.5) 1 mM ethylenediaminetetraacetic acid (pH to 8.0 to dissolve) 0.1 N sodium hydroxide 0.5 % sodium dodecyl sulfate2.3 M Sodium acetate, pH 5.2 3.Pre-ch[阅读全文]

摘要:1.grow up single colony in 1.5 ml LB/antibiotic ovn @37℃ .2.pour into tube, spin for 30 sec .3.decant supernatant and resuspend pellet in 100 ml lysis solution .4.add 200 ml alkaline SDS, vortex well [阅读全文]

摘要:There are many methods for DNA preparation without using kits.Below are a few minipreps.These protocols may be scale up to midi or maxi prep if necessary.1.Ammonium Acetate Method 1)Spin down cell and[阅读全文]

摘要:1. Spin down 1.5 ml of overnight culture in 2ml or 1.5ml tube for 1 minute on high. 2. Asprirate supernatant and resuspend cell pellet in 100 µl Solution I (25 mM Tris-HCl, pH 8.0, 10 mM EDTA). [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:This protocol is for Mini (up to 20 µg) preparations of high-copy plasmid DNA from cultures of E. coli .Important notes before startingNew users are strongly recommended to read Appendix A (page[阅读全文]

摘要:Reagents:Soln 1: 50 mM glucose/10 mM EDTA/25 mM Tris pH 8. Autoclave before use. Add 2 mg/ml Lysozyme just prior to use.Soln 2: 0.2 N NaOH/1% SDS. Keep solution at room temperature. Solution is normal[阅读全文]

摘要:Important Notes Before Starting • New users are strongly recommended to read the QIAGEN handbook before starting the procedure. •Before using the kit for the first time, dissolve the lyophil[阅读全文]

摘要:Purpose:The method is used to clone smaller portions of inserts (up to 15 kb in size)which previously have been cloned in YACs phage cosmids or other plasmids.Time required:Plasmid Vector PreparationR[阅读全文]

摘要: Materials:TE buffer10% (w/v) sodium dodecyl sulfate (SDS)20 mg/ml proteinase Kphenol\chloroform (50:50)isopropanol70% ethanol3M sodium acetate pH 5.2Phase Lock GelTM (5 Prime, 3 Prime, Inc)1.Grow E. [阅读全文]