Alkalineagarosegelelectrophoresis
Alkaline agarose gel electrophoresis (Sambrook et al., 1989)Alkaline agarose gels can be used to determine the size and quality of first and second strand cDNA synthesis by reverse transciptase.You will need:Alkaline loading buffer (300mM NaOH, 6mM EDTA, 18% Ficoll 400, 0.15% bromocresol green, 0.25% xylene cyanol FF)10 x Alkaline agarose electrophoresis buffer (500mM NaOH, 10mM EDTA, pH 8.0, made fresh for each use)......阅读全文
细胞支原体污染的PCR检测
支原体菌株来源：M.Arginini ATCC23838 精氨酸支原体M.FermentaneATCC19989 发酵支原体M.SalivariumATCC23064唾液支原体M.HominisATCC23114人型支原体M.OraleATCC23714口腔支原体M.HyorhinisATCC290
细胞支原体污染的PCR检测
支原体菌株来源： M.Arginini ATCC23838 精氨酸支原体 M.FermentaneATCC19989发酵支原体 M.SalivariumATCC23064唾液支原体 M.HominisATCC23114人型支原体 M.Ora
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
DNA克隆
DNA克隆（主要内容如下）· General Procedure· PCR Cloning· Subcloning· ET Cloning· Vector Preparation· Ligation Re
ALKALINE－PHOSPHATASE－(APAAP)－TECHNIQUE
Preparation: Cytological PreparationsFixation: Air dry films or cytospin preparations overnight at room temperature. For frozen and paraffin sections
Isoelectric－Focussing－of－Membrane－Proteins－by－Slab－Gel－Method
REFERENCE: Ames, G.F.L. and Nikaido, H. 1976. Biochemistry. 15:616-623.MATERIALS:Gel solution:1.05 gacrylamide0.032 gbis-acrylamide8.25 gurea6.5 mldis
Phage－DNA
IntroductionThe phage lysate from the plate contains bacterial DNA and RNA, as well as phage DNA encased in the phage coat. The following procedure, d
基于PCR技术的染色质沉淀分析
INTRODUCTIONAfter chromatin immunoprecipitation (ChIP), different PCR-based approaches can be used to determine how much DNA is precipitated at a locu
甲醛洋菜胶体电泳
实验概要本实验介绍了甲醛洋菜胶体电泳 (formaldehyde-agarose gel electrophoresis)的操作流程。实验原理甲醛是一种常用的RNA 变性剂，进行电泳时所使用的缓冲液为MOPS (3-[N-morpholino] propanesulfonic acid)。由于r
Competitive－RTPCR－Strategy－for－Quantitative－Evaluation－4
We have also been able to detect expression of this receptor in all studied tissues, which is consistent with the pleiotropic nature of growth hormone
TRFLP的优缺点
该技术在应用的过程中，肯定需要在实验条件上不断进行改进，而这种改进的好坏自然而然需要实验结果的验证。。V. Grüntzig在2002年做了该工作的一部分，结果认为，在限制性酶切时，很有必要去除其中影响DNA的酶切过程，并且实验证明了具体的酶切时间。具有说服力的结果如下： 1，T-RFLP出数据的
TRFLP技术的优缺点
T-RFLP（末端限制性片段长度多态性）该技术在应用的过程中，肯定需要在实验条件上不断进行改进，而这种改进的好坏自然而然需要实验结果的验证。V. Grüntzig在2002年做了该工作的一部分，结果认为，在限制性酶切时，很有必要去除其中影响DNA的酶切过程，并且实验证明了具体的酶切时间。具有说服力的
甲醛洋菜胶体电泳实验
甲醛洋菜胶体电泳 （formaldehyde-agarose gel electrophoresis）甲醛是一种常用的RNA 变性剂。用于（1）RNA的分离测定（2）RNA提纯。实验方法原理rRNA 占细胞RNA总量的80~85%,以ethidium bromide 染色后，呈现于胶体上的两个主要R
酵母人工染色体
· Easy YAC Preparation Method (Andrew Davies,Shaw lab)· Screening YAC libraries (Donis Keller Lab)This is a method for screening YAC l
DNA－labeling－by－nick－translation
DNA labeling by nick translationreagents: DNA for labeling (concentration c > 150 ng/µl) modified nucleotides: Biotin-16-dUTP, Digoxigenin-11-dUTP, co
Large－Scale－Plasmid,－Cosmid,－BAC,－PAC,－and－Fosmid－DNA－Isolation
DNA Isolation by a Cleared Lysate Method Followed by Double Acetate Precipitation Version 3b - updated September 26, 1999The Most Recent Roe Lab Imple
Thermal－Inactivation
Thermal InactivationA simple, reversible way to a stop restriction reaction is by adding EDTA, which chelates Mg2+, thereby preventing catalysis. If f
Human－Placental－Alkaline－Phosphatase－Stain
The following protocol is for detection of human placental alkaline phosphatase (AP) in cultured cells. Human placental AP is heat stable, unlike othe
Apoptosis:－A－Laboratory－Manual－of－Experimental－Methods－Andrea－Cossarizza
THE CELL 1. Morphological aspects of apoptosis Walter Malorni, Stefano Fais & Carla Fiorentini 2. Cell cycle Miriam Capri & Daniela BarbieriTHE NUCLEU
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
Construction－of－BAC－Libraries：Megabase－DNA－Isolation
Megabase DNA IsolationMegabase-size DNA isolation from plantsTo construct large insert DNA libraries in BAC and YAC vectors, methods must be developed
Thermal－Cycling－Profile－for－Standard－PCR
Initial denaturationIt is very important to denature the template DNA completely. Initial heating of the PCR mixture for 2 minutes at 94°–95°C is enou
Southen杂交
Southern杂交One important thing for transfer:the weight of the object resting on top of the blotting apparatus should not exceed the weight equal to a 5
An－Integrative－Procedure－for－Apoptosis－Identification－and－Measurement2
TroubleshootingCritical Steps(1) Don’t trypsinize cells for too long when collecting them.(2) Rotation speed should be no more than 1500 rpm during ce
DNA转化实验指导2
1B. Cloning 1. A caveat on dephosphorylation: the most common reason for failure to obtain colonies is a result of adding too much BAP or CIP to
electrophoresis－of－DNA
Agarose Gel Electroporesis of DNA Making the gel: 1. Place casting platform with well former sideways in gel stand where you wish to pour
Chromatin－Electrophoresis
Dr. William H. Heidcamp, Biology Department, Gustavus Adolphus CollegeExercise 10.4 - Chromatin ElectrophoresisLEVEL IIMaterials 14 M Urea6 M NaCl0.05
Protein－Electrophoresis
DefinitionAmino acids, nucleotides, polypeptides, and other compounds in a colloidal state can be separated by the application of external voltages wh
Capillary－Electrophoresis
Capillary electrophoresis is a very sensitive analytical technique. Sample components are separated within a fused silica capillary using one of sever
DNA转化实验指导5
2C. Notes on Transformation 1. Bacto tryptone and yeast extract can cause allergic reactions. 2. All containers used to handle the bacteria (