摘要:Protocol:Combine an appropriate concentration of probe that has been labeled on one strand (100-150 fmol probe has worked so far) with the appropriate concentration of protein in a total volume of 50&[阅读全文]

摘要:Recovery of DNA from Low Melting Point Agarose Gels1.Run digestion products on 0.7% LMP agarose gel in 1X TBE (it's nice to have at least 1ug of the fragment you want). LMP agarose is fragile; pour ge[阅读全文]

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摘要:第一节 概 述 一. DNA的限制性内切酶酶切分析 限制性内切酶能特异地结合于一段被称为限制性酶识别序列的DNA序列之内或其附近的特异位点上，并切割双链DNA。它可分为三类:Ⅰ类和Ⅲ类酶在同一蛋白质分子中兼有切割和修饰([阅读全文]

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摘要:DNaseI FootprintintSolutions10X Binding Buffer200 mM Tris 8.0 200 m l 1M Tris pH 8.0500 mM NaCl 100 m l 5M NaCl10 mM EDTA 20 m l 0.5 M EDTA pH 8.0680 m l Qstore at room temperatureDNaseI Dilution Buff[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: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]

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摘要:This is a modification of a salting out procedure as described by Miller et al., (1988), evaluated at the DNA Laboratory, Medical School, Malta. When analysed by spectrophotometry >95% of extracted ge[阅读全文]

摘要:Restriction digests consist of:15.75 ml ddH2 O 1 µl 10 X buffer B (Boehringer Mannheim) 0.25 µl HindIII (40 U/µl) 3 µl DNA Set up digestions in 96 well plates. Incubate at 37℃ [阅读全文]

摘要:Pouring the Gel Outline: Pouring this big & thin 6% acrylamide gel ("Mother of all gels") is quite a challenge and probably the reason why smart whimps by them ready to use.Supplies & Equipment: water[阅读全文]

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摘要:1.Obtain 65-100 µl of blood by retro-orbital bleed with a heparinized microcapillary tube. Expel blood immediately into a 1.5 ml microfuge tube containing 20 µl of 10 mM EDTA. Mix immediat[阅读全文]

摘要:1. Remove 0.5 cm of tail into polypropylene microfuge tube (do not mince).(The tubes must have tight-fitting caps, so that there are no leaks in steps 3 and 7 below.)2. Add 0.5 ml DNA digestion buffer[阅读全文]

摘要:Phenol-chloroform DNA extraction from sand flies1. Crash Individual sand fliesin 0.5 ml lysis buffer (50 mM NaCl , 10 mM EDTA, 50 mM Tris-HCl pH 8). 2.Incubate The sand fly homogenateswith 100 ng/ml R[阅读全文]

摘要:1. Remove gel slice contain DNA fragment and place in 10 volumes of:300 mM NaOAc, pH 7.0 300 ml 1 M NaOAC, pH 7.01 mM EDTA 2 ml 500 mM EDTA, pH 8.0698 ml ddH2O2. Incubate at 22 ℃ for 30 min. Transfer [阅读全文]

摘要:序列测定的技术和策略Sanger双脱氧链终止法Maxam－Gilbert DNA 化学降解法测序策略目前应用的两种快速序列测定技术是Sanger等（1977）提出的酶法及Maxam和Gilbert(1977)提出的化学降解法。虽然其原理大相径庭，但这两种方[阅读全文]

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