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摘要:20cm Gel Preparation, Sample Separation, and Visualization: Glass Plate Preparation:[阅读全文]
摘要:Electroelution of agarose fragmentsElectroelution buffer1 M Tris, pH 7.5 12.0 mls0.5 M EDTA 0.24 mls1 M NaCl 3.0 mlsqs to 600 mls dH2OAcetate cushion 3 M NaAcetate pH 4.8 480 μl0.1 % Bromphenol Blu[阅读全文]
摘要:PrincipleThe nucleic acids contained within the gel slice are electrophoresed out of the gel, funneled down into the v-shaped channel and trapped within a high salt cushion resting at the bottom of th[阅读全文]
摘要:PrincipleThe nucleic acids contained within the gel slice are electrophoresed out of the gel, funneled down into the v-shaped channel and trapped within a high salt cushion resting at the bottom of th[阅读全文]
摘要:Amberg Lab ,Upstate Medical University http://www.upstate.edu/biochem/amberg/protocols/transformation.html1.Grow cells to 1X10E8 or OD600 of 1.2-1.3. 2.Spin cells at 5,000rpm for 5 min, and wash pelle[阅读全文]
摘要:Sugden lab,McArdle Laboratory for Cancer Research,University of Wisconsin-Madison Medical School DNA /EMSA.pdf">http://mcardle.oncology.wisc.edu/sugden/Protocols/PDF files/Association of protein with [阅读全文]
摘要:Hahn Lab，The Fred Hutchinson Cancer Research Center and Howard Hughes Medical Institute A -80 degree C frozen stock is prepared from Gibco-BRL DH10B competent cells.From this frozen stock,a freshly st[阅读全文]
摘要:1.O/N (5ml E.coli in LB-->1ml into 200ml LB/37℃2.Grow 2-3H to A660 ~0.3-0.43.Chill cells 10'/ice Spin down 10',5K,4℃4.Wash 1X 200ml 10% glycerol (sterile)40℃; Spin down 10',5K,4℃5.Wash 1X 40 ml 10% gl[阅读全文]
摘要:The manipulation of embryonic stem (ES) cells to generate targeted mutations via homologous recombination has proved an invaluable resource for researchers from fields as diverse as embryology, immuno[阅读全文]
摘要:Cells are routinely passaged two days prior to electroporating. Usually one 10 cm plate at approximately 80% confluency will provide enough cells for 1-2 electroporations. Procedure 1. Change medium o[阅读全文]
摘要:Need 1.5-2 x 107 cells from a 2 day culture. 1. Cells are harvested as normal, washed x 1 in PBS then taken up at conc. of 1.2 x 10 7 cells/ml in cold PBS. 2. Take x2 Biorad curvettes A. 0.9 ml (107 c[阅读全文]