NuPAGEGels
NuPAGE GelsA gel electrophoresis system used for SDS-PAGE protein analysis. The gels are made up of Bis-Tris-HCl (pH 6.4) polyacrylamide and are intended for denaturing conditions only.NuPAGE Electrophoresis Protocols1) Remove gel from pouch and rinse gel with D.I. water holding the gel by the edges of the cassette. (Gels should be stored at 4º C)2) Mark the bottom of the lanes with a permanent marker.3) Peel off tap......阅读全文
Basic－Protein－Chemistry－Techniques
Coomassie Blue Stain: (for gels) 1) Combine 225 ml Methanol with 225 ml ddH2O. 2) Add 0.5 grams of Coomassie Blue. 3) Just before use, add 50 ml acet
Cloning－of－small－RNAs－with－5’－phosphate－and－3’－OH－ends2
3’ Adaptor Ligation and PurificationHeat shock the RNA by putting at 90°C for 30 seconds. Snap cool on ice.Set up the 3’Adaptor ligation reaction in a
Methylene－Blue－DNA－staining－protocol
Methylene Blue DNA staining protocolProtocol:Load 2-5X the amount of DNA that would give bands of moderate intensity on an ethidium bromide stained ge
Basic－Protein－Chemistry－Techniques
实验概要Basic Protein Chemistry Techniques实验步骤Coomassie Blue Stain: (for gels) 1) Combine 225 ml Methanol with 225 ml ddH2O. 2) Add 0.5 grams of Coomassi
Assay－of－superoxide－dismutase－activity1
Assay of superoxide dismutase activity by combining electrophoresis and densitometryAbstract. A modified technique was developed to assay superoxide d
Assay－of－superoxide－dismutase－activity2
Cuvette holders in the sample chamber of the spectrophotometer were thermo-controlled at 25°C. For the blank test, 100 ml of 50 mM potassium phosphate
Agarose－Gel－Electrophoresis
实验概要Separating nucleic acid fragments by agarose gel electrophoresis.实验原理 Agarose gel electrophoresis remains the most widely used technique for sep
Lipoprotein－Analysis－Week－2:－Electrophoresis
Lipoprotein Analysis Week 2: Electrophoresis IntroductionSDS polyacrylamide gel electrophoresis (SDS PAGE) will be used to assess the purification pr
非变性胶蛋白电泳
Section 2.1Nondenaturing Polyacrylamide Gel Electrophoresis of ProteinsJohn M. Walker1. IntroductionSDS-PAGE (Section 2.2) is probably the most commo
BioRad（伯乐）Western－Blot半干法转膜的十大注意事项
首先讲转膜仪的清洗：Do not immerse the unit in liquid. Use special care when cleaning the anode plate to avoid scratching or marring the platinum. Do not use
Total－Protein－Extraction－with－TCAAcetone
We describe a procedure allowing extraction of total proteins that performs efficiently with a large variety of plant tissues, based on simultaneo
General－Cloning－Protocols
Large Scale Preps: (See Large scale plsasmid prep protocol for more details)Cultures: Inoculate a 5 mL LB/Amp (50 - 100 µg/mL) culture in early a.m. w
ISOLATION－OF－RNA－FROM－BACTEROIDS
3 g nodules (fresh or frozen in liquid N2) were ground to a powder in mortar and pestle with liquid N2. To the powder was added ice cold 0.5 M mannito
酵母准备
Yeast DNA PreparationYeast Genomic Preparation (Gottschling Lab)Rapid method for yeast genomic DNA isolation Yeast DNA Preparation (rapid glass bead
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
Native－gel－electrophoresis（非变性电泳）
Native gel electrophoresis Under native PAGE conditions, polypeptides retain their higher-order structure and often retain enzymatic activity and inte
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
Cloning－of－small－RNAs－with－5’－phosphate－and－3’－OH－ends3
Load your precipitated PCR samples into 2 consecutive lanes so as not to overload the lanes. For each different sample, I would run a separate ladder
Ingel－digestion－of－proteins－for－peptide－fingerprint－mapping
Polyacrylamide gel electrophoresis is a widely used technique to separate proteins from biological samples. Moreover, the development of two-dimension
peptide－fingerprint－mapping
Polyacrylamide gel electrophoresis is a widely used technique to separate proteins from biological samples. Moreover, the development of two-dimension
2D－Polyacrylamide－Gel－Electrophoresis
This method was successful in our lab using prostate tissue and for our specific objectives. Investigators must be aware that they will need to tailor
DNA抽提
DNA抽提（主要内容如下）· Working with DNA· DNA Extraction from Bacteria and Other Organisms· DNA Extraction from Cell and Tissue· Mitochondria DNA Isola
PIC－Crosslinking－and－Immune－Precipitation
References Fishburn et. al., 2005, Molecular Cell, vol. 18 #3: Experimental Procedures pg. 376Immobilized Template assay (Hahn lab website)NotesDTT mu
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
寡核苷酸的相关操作
In this section, you will find techniques related to oligonucleotides, such as oligo purification by acrylamide gel, annealing two oligos to make doub
SOUTHERN－BLOT的步骤
1. Run the gel as normal. Often for genomic southerns it is desirable to run long gels (18cm) over 4-6hrs.2. Photograph the gel with a ruler adjacent
Southern杂交技术
SOUTHERN BLOT1. Run the gel as normal. Often for genomic southerns it is desirable to run long gels (18cm) over 4-6hrs.2. Photograph the gel with a r
即时可用的蛋白质电泳标准
Choose a protein standard MultiMark®SeeBlue® Plus2/SeeBlue®BenchMark™ Pre-stainedMark™BenchMark™MagicMark™ XP newIEF mARKER 3-10ApplicationSDS-PAGEGoo
PCR－clean－up
Following PCR, you often want to get rid of the PCR primers and Taq polymerase before the next step. This is necessary for sequencing PCR products or
High－Resolution－Agarose－Gel－Electrophoresis
实验概要Agarose gel electrophoresis remains the most widely used technique for separating nucleic acid fragments due to its ease of use, non-toxicity, a