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RNA提取的基本操作步骤
RNA的提取准备试剂:氯仿,异丙醇,75%乙醇,无RNase的水或0.5%SDS(溶液均需用DEPC处理过的水配制) 操作步骤: 1. 匀浆处理: ①
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总RNA的提取(Trizol法提取)
在收集到生物材料之后,最好能即可进行RNA制备工作。若需暂时储存,则应以液氮将生物材料急速冷冻后,储存于-80℃冷冻柜。在制备RNA时,将储存于冷冻柜的材料取出
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动物组织RNA的提取
实验目的:掌握由动物组织中提取总RNA的方法 实验原理:Trizol试剂是一个包含酚、异硫氰酸胍和SDS的单相酸性溶液,其在裂解细胞的同时抑制RNase的活
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"Dirty" Mini Preps
1) Grow 3-5 ml over-night E. coli cultures containing your plasmid 2)
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Thermal Inactivation
A simple, reversible way to a stop restriction reaction is by adding EDTA, which
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Partial Endonuclease Digestion
Prepare a 100 µl reaction mixture containing the DNA of interest in the appropri
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EXO/S1 DELETION SERIES
1. Cut DNA with 5'- and 3'- overhangs, gel check, phenol-Sevag extract
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NEB识别序列
下表中列出了NEB提供的所有限制性内切酶的特异性识别序列,同时列出的还有回文序列的每条互补链。比如CCTC(7/6) 和(6/7)GAGG均显示为MnlI识别序
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Restriction Digestion of DNA
Restriction enzymes are expensive ($40 to $200 a vial): keep the enzymes at -20℃
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UV Shadowing
UV shadowing is a technique for visualizing nucleic acids separated on acrylamid
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FOOTPRINTING WITH DNASE1
1)probe: same as used for gelshift), isolated by isotacelectrophoresis w/out EtO
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DNA rapid CTAB
A Rapid CTAB DNA Isolation TechniqueUseful for RAPD Fingerprinting and Other PCR
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连接互相匹配的粘性末端,产生新的酶切位点
通常连接互相匹配的粘性末端可以产生新的酶切位点。以下组合是通过计算机程序计算出来的,尽管我们希望能尽量保证其准确性,但有些未经实验证实,因此不能保证100%的可
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创造新的酶切位点
将互相匹配的DNA末端连接起来可以产生新的酶切位点。这些DNA末端可以通过以下方法获得: 1. 补平5′突出的粘性末端产生平末端 2. 双酶切产生的平末端 3.
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Restriction digestion
Restriction enzyme digestions are performed by incubating double-stranded DNA mo
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