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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 ...

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

This method is after MarchukD.et al.1991Nucl.Acids Res.19(5)pp1154. You will need:10 x Taq buffer (Promega)Taq Polymerase (Promega)Phenol/chloroform mix100mM dTTPTE bufferAbsolute ethanol70% ethanol 1 ...

Hattoti's protocol adapted to cell culture :Hattori MTugores AVeloz LKarin MBrenner D (1990)A simplified method for the preparation of transcrip-tionally active liver nuclear extracts.DNA Cell Biol.Vo ...

1) The ligation mixture contains the following:vector (~100 ng)　　insert (equimolar or 2 or 3 X molar concentration of vector)　　water added to 18 µl2) Heat the mixture at 45 ℃ for 5 min. to melt any an ...

NH4Ac and EtOH precipitation of DNA Add NH4Ac (10M stock or solid) to the sample for a final concentration of 2.5M mix (spin at 4℃ transfer the supernatant to a new tube; optional spin for extra purif ...

PROTOCOL TO EXTRACT DNA FROM PARAFFIN BLOCKS1.Cut 10-20X10μm sections of formalin fixed paraffin samples into eppendorf tubes.2.Add 1 ml xylene mix incubate at 55℃ for 15mins. Release pressure spin ...

CTAB TECHNIQUE / Method / Schedule / Protocol FOR DNA ISOLATION / DNA EXTRACTION FROM PLANT LEAF / LEAVES SAMPLES(see also DNA RNA double isolation procedure if both DNA and RNA are needed)Reagents ne ...