ExposinggelsandplatescontainingradioactivesamplestoXrayfilm
Although most people use the PhosphorImager for western blots, kinase assays and methionine-labeled samples, X-ray film remains the best way to expose peptide maps and prep gels. TerminologyAutoradiography is the direct exposure of film by beta particles or gamma rays. Fluorography is the exposure of film by secondary light that was generated by the excitation of a fluor or a screen by a beta part......阅读全文
Exposing－gels－and－plates－containing－radioactive－samples－to－Xray－film
Although most people use the PhosphorImager for western blots, kinase assays and methionine-labeled samples, X-ray film remains the best way to expose
PREPARATION－OF－SEQUENCING－GELS
MATERIALS:2-glass plates1 sharks -tooth comb and spacersWhatman 3 mm paper30 or 40% acrylamide-bis (19:1)10X TBEurea10% ammonium persulfateTEMED60 cc.
Polyacrylamide－Gel－Electrophoresis－of－Oligonucleotides
1. Pour and polymerize a 20% polyacrylamide gel, no Urea.2. Remove clamps. Rinse with water. Remove comb. Rinse top of gel well.3. Insert comb teeth d
基于PCR技术的染色质沉淀分析
INTRODUCTION After chromatin immunoprecipitation (ChIP), different PCR-based approaches can be used to determine how much DNA is precipitated at a loc
Marcantonio－Lab－Protocol－Manual——3
Sequencing GelPreparing and Running a Sequencing Gel (6% Polyacrylamide/Urea) A. Preparation of Gel Solution 1) Weigh out 50 g of Urea into a clean 25
基于PCR技术的染色质沉淀分析2
METHOD Analysis of precipitated chromatin fractions (from Chromatin Immunoprecipitation on Unfixed Chromatin from Cells and Tissues to Analyze Histone
Analysis－of－Proteins－using－Small－Format－2D－Gel－Electrophoresis
Preparation of protein samplesIntracellular virus proteinsThe following method has been developed principally for the analysis of intracellular protei
SSR－GEL－and－Silver－Staining－Protocol
I. EQUIPMENT:DNA sequencing unit (35 x 45 cm) & 2000V power supplyClampsLg. plastic trays (4), about 43 x 50 x 8 cm, and one lidTwo rocking platformsH
DGK－Assay
Buffers:- 2X buffer10 ml 0.5 M imidazol, pH 6.60.21 g LiCl1.25 ml 1 M MgCl21.0 ml 0.1 M EGTA, pH 6.6--> Bring volume up to 50 ml with distilled water.
Denaturing－Gradient－Gel－Electrophoresis－(DGGE)
Purpose:Denaturing gradient gels are used to detect non-RFLP polymorphisms. The small (200-700 bp) genomic restriction fragments are run on a low to h
AA－－Metabolite－Quantitation－of－Media－PostAA－Labeling
1) Remove 2 500 µl aliquots of supernatant into scintillation vials, add scintillation fluid and count.2) Aliquot 1.6 mls of the remaining supernatant
AA－－Metabolite－Quantitation－of－Cell－Pellets－PostAA－Labeling
Extraction:1) Following spin, save supernatant for analysis. Be extremely careful not to disturb the pellet since it is somewhat dispersed.2) Immediat
Genomic－Libraries
Genomic DNA libraries Size of some genomes and chromosomes:Comparative Sequence Sizes(Bases)(yeast chromosome 3)350 ThousandEscherichia coli (bacteriu
基于PCR技术的染色质沉淀分析1
INTRODUCTIONAfter chromatin immunoprecipitation (ChIP), different PCR-based approaches can be used to determine how much DNA is precipitated at a locu
Phosphoamino－acid－analysis:Mark－Kampss－method
Phosphoamino acid analysis:Mark Kamps's method1. Label your protein with 32Pi. Then subject the protein to SDS polyacrylamide gel electrophoresis
Assay－of－superoxide－dismutase－activity1
Assay of superoxide dismutase activity by combining electrophoresis and densitometryAbstract. A modified technique was developed to assay superoxide d
Hybridization－of－High－Density－Arrayed－BAC－Nylon－Filter－Blots
Protocol for hybridization of high density arrayed Bacterial Artificial Chromosome nylon filter blots with 100 PCR isolated Unigene cDNA inserts, pool
CarbohydrateSpecific－Adhesion－of－Intact－Cells－to－Resolved－Glycolipids－on－T
Carbohydrate-Specific Adhesion of Intact Cells to Resolved Glycolipids on TLC PlatesRonald L. Schnaar~Professor, Johns Hopkins University Medical
UV－CrossLinking－an...－（二）
实验步骤1. UV cross-linking of tissue culture cells 1) Remove media and add ice-cold PBS to cells (e.g. use cells grown in a 10 cm plate for three ex
Assay－of－superoxide－dismutase－activity3
Botanical Bulletin of Academia Sinica, Vol. 37, 1996The method using NBT as a superoxide radical competitor and a color indicator was also explored to
蛋白质检测
· Protein detection (Aberdeen's Lab)The method used to locate the proteins following 2D-PAGE depends on the nature of the original sample.
重组DNA的分离、克隆与测序实验手册2
C. Restriction digestionRestriction enzyme digestions are performed by incubating double-stranded DNA molecules with an appropriate amount of restrict
A－Method－for－Assaying－Deubiquitinating－Enzymes2
Table 1: Hydrolysis of 125I-labeled Ub-PESTc by the purified YUH1.Specific activity againstDUBsCbz-LRGG-AMC125I-labeled Ub-PESTcYUH13.2 x 10-105.1 x 1
Methods－for－the－Measurement－of－a－Bacterial－Enzyme－Activity－in－Cell－Lysates2
Measurement of ACTase activityNuclear magnetic resonance spect roscopy (NMR)The unique potential of NMR spectroscopy for monitoring simultaneously the
Amino－acid－composition
There has been a recent revival of interest in the use of AA composition for the identification of proteins from 2-D gels. This technique uses the idi
ELECTROPHORESIS－OF－DNA－IN－AGAROSE－GELS
ELECTROPHORESIS OF DNA IN AGAROSE GELSA). AGAROSE CONCENTRATIONS: Use 0.8% agarose (w/v) for high molecular weight DNA fragments, and 1 - 1.2% f
Detection－and－Measurement－of－Radioactivity
Radioactive DecayIsotopes of a given element have nuclei with the same number of protons but different numbers of neutrons. Some isotopes are stable,
Pouring－Plates
1. For rich media, weigh out appropriate ingredients and place into a flask. Add water until appropriate volume. Use a flask at least 2 times larger t
Layered－plates
General InformationThe cells being plated are sandwiched between layers of cell-free agar. The submerged cells grow into smaller colonies and many mor
Exercise－12.10－－Establishment－of－a－Primary－Culture
Exercise 12.10 - Establishment of a Primary CultureLEVEL IIIMaterialsChick embryo (approximately 8 days old)70% (v/v) ethanol for swabbingSterile scis