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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 ...

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 ( ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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1)Prior to any ligation reactionyou should always run one gel in which purified insert and cut backbone are run side by sidepreferably beside a known amount of cut DNA.You can then use this to estimat ...