摘要:1. Add 400 μl of TE buffer then 400 μl of 1-butanol to the oligonucleotide glass vial.2. Vortex well, then spin down in tabletop centrifuge for 10 minutes at ~2,000. rpm¹s.3. Remove top, bu[阅读全文:]
摘要:This protocol was designed to generate directionally end-labeled probes for DNaseI footprinting but it can be used for any application that requires end-labeled DNA probes.Solutions10 mM dNTP StocksTh[阅读全文:]
摘要:Glass wool method: Run TAE agarose gel and cut the appropriate band out with a clean razor blade. Poke a small hole with hot needle on the bottom of an eppendorf tube, and jam the hole with a little o[阅读全文:]
摘要:TE solution 10 mM Tris (pH to 7.5) 1 mM EDTA (pH to 8.0 to dissolve)2.Frozen agarose gel piece containing the desired DNA fragmentSupplies: 1.Micropipetter and tips 2.Microcentrifuge and tubes 3.Spatu[阅读全文:]
摘要:Sephadex G-50 spun column purification Spin column purification can be used to change buffers without a concomitant change in solution volume, to remove protein contaminants or to purify plasmid DNA s[阅读全文:]
摘要:1. Pick single colony and inoculate 250 ml of LB broth containing 100 m g/l ampicillin or appropriate antibiotic. Shake at 250 RPM overnight.2. Centrifuge cells in a Sovall GSA (250 ml)or SLA-3000 (50[阅读全文:]
摘要:This protocol works well from a 5 ml starting culture or a 1 ml starting culture (adjust volume accordingly).Solutions2X YT Media16 g tryptone10 g yeast extract5 g NaCl1 ml 1N NaOHup to 1 liter with Q[阅读全文:]
摘要:Materials: RPMI 1640 medium fetal calf serum (FCS), 20% Colcemid (e.g. Boehringer Mannheim cell biology reagents, Best.-Nr. 295892) cell cuture flask Phythemaglutinin, PHA-L (Seromed, M 5030) CO2 cell[阅读全文:]
摘要:SolutionsGel StocksDiluent 5X Buffer 25% Acrylamide209 g Urea 209 g Urea 209 g Ureaup to 500 ml Q 250 ml 10X TBE 120.8 g Acrylamideup to 500 ml Q 4.1 g BISup to 500 ml Q2.5 M NH4 OAc19.2 g NH4 OAcup t[阅读全文:]
摘要:This is a very fast mini-prep protocol which is suitable for sequence analysis and restriction digests. Although the yield is higher than Protocol D.1, there is considerable chromosomal DNA, RNA and p[阅读全文:]