DNA实验技术

Non-radioactive Probes I. Via random hexamers 1. Solutions:10X hexa nt miundefined: 500 mM Tris-Cl pH 7.2100 mM MgCl21 mM dithioerythritol (DTE)2 mg/ml BSA62.5 A260 units/ml (1.56 mg/ml) random hexanucleotid ...

RNA/Zeta Probe Dot Blotting protocol1)Make up RNA (up to 20μg)dissolved in sterile H2 OTE or 0.5% SDS to 500μl with ice-cold sterile 10mM NaOH1mM EDTA and apply it to Zeta Probe membraneheld in ...

The standard solution typically used for both pre-hybridisation and hybridisation is based on that given in Maniatis et al.(1982)with both Denhart's solution and heterologous DNA being replaced by hep ...

About 1/2 to 1 cm of tail should be cut from properly marked mice (using toe or ear clipping etc) about the age of 2-3 weeks. Place these tails into marked 1.5 ml Eppindorf tubes for processing; they ...

1. Depurination of DNA fragments: Wash gel in 0.25 M HCl 5' (small gels) 7' (large gels)2. Denaturation: Wash gel in (0.5 M NaOH 1.5 M NaCl) for 20' (small) to 30' (large).3. Neutralization: Wash in ( ...

一 酶切和电泳在200 μl 微量离心管中加入：25 μl DNA样品(约10μg)，3μl 限制性内切酶（MBI，10 U/ μl）5 μl 相应的10×buffer，补水到50μl。然后加一滴矿物油覆盖 稍微离心后放于37℃水浴8-12小时。酶切完后，取5μl 酶切DNA样品于0.8%的琼脂糖凝胶上检测酶是否充分。如果酶切充分，灌制0.8%的ag ...

For use with GibcoBRL Random Primer DNA labeling system. Objective: To produce radioactively labeled DNA strands for the detection of target DNA or RNA sequences in various applications including Sout ...

1) 取10μl待测DNA，于一定浓度的琼脂糖凝胶上进行电泳。(20mL，1.0%凝胶)2) 凝胶用溴化乙锭染色，切掉凝胶四周多余部分，并在凝胶的一角作一记号， 拍照记录电泳结果(拍照时， 凝胶旁放一尺子)。3) 杂交用胶的制备 (做以下步骤两组合一)A. 将凝胶浸没入30mL 0.25mol/L HCl溶液中15min，使DNA脱嘌呤。B.用蒸馏水短暂洗凝胶2次。C.将凝胶浸没入30mL变 ...

Analys of Genomic DNA by Southern Hybridization (Southern Blot) Outline: Localization of particular sequences within genomic DNA is usually accomplished by the transfer techniques described by Souther ...

reagents: DNA for labeling (concentration c 150 ng/µl) modified nucleotides: Biotin-16-dUTP Digoxigenin-11-dUTP conc. 1nmol/µl (Boehringer Mannheim) dNTPs (regular nucleotides): dATP dCTP dGTP 0.5 mM ...

SolutionsProcedure Design complementary oligonucleotides with compatible half sites on the ends (I use BamHI and BglII). Dry down 300 ng of the two purified oligos together and resuspend in 9 m l Q wi ...

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 ...

Wear gloves throughout and work in radiation area. Monitor area before and after use.Mix the following in an eppendorf tube:1. 0.5 microgram oligonucleotide dissolved in H2O.2. 3 microliters 10x kinas ...

The gel shift or electrophoretic mobility shift assay provides a simple and rapid method for detecting DNA -binding proteins. This method has been used widely in the study of sequence-specific DNA -bi ...

1. Prepare Whatman DE-52 according to manufacturers specifications. Equilibrate at store with 0.02% Sodium azide.2. Plug a 1 mL (blue) pipet tip with Siliconized glass wool. Add 500 ml of 50:50 DE-52 ...

Protocol for Annealing OligonucleotidesOligo Name:Lot Number:Total nmol:Volume of Annealing Buffer added:Oligo Name:Lot Number:Total nmol:Volume of Annealing Buffer added:Annealing Buffer: 10mM Tris p ...

In the hood:96 well dish with bacteriatitertechmicrotubesglass pipetteRemove colonies from each well using the titertech and place them into the coverPipette up and down to thoroughly mix the colonies ...

染色质免疫沉淀法（Chromatin immunoprecitation，ChIP）是研究体内DNA与蛋白质相互作用的重要工具。它可以灵敏地检测目标蛋白与特异DNA片段的结合情况，还可以用来研究组蛋白与基因表达的关系。核小体组蛋白可以发生多种翻译后的共价修饰，如乙酰化、甲基化、磷酸化、泛素化等，这些共价修饰与真核基因的表达密切相关。根据“组蛋白密码”假说，组蛋白的各种共价修饰的组合会以协同或拮抗的 ...

PurposeThe protocol describes how to prepare human tonsil lysate for use in purification of ICAM-1LFA-1and PNAd.MaterialsSafety EquipmentsLab CoatLatex GlovesFace MaskBench PaperFresh human tonsil tis ...

Does anybody know how to isolate single-stranded DNA from mammalian cells?Thanks for responding.Yin----------------------------------------------------------------------------------HelloI don't know i ...