RestrictionDigestsofHighMolecularWeightYeastDNA
Purpose:To perform restriction digests of YACs for mapping using rare cutting enzymes or more conventional restriction endonuclease digestion.Time required:2-4 hoursSpecial Reagents:dependent upon enzymes desiredProcedure:Using wide-bore pipette, aliquot 100 ng of liquid DNA into eppendorf tube. Add 2.5 祃 10X restriction buffer, 1-2 ul RNase (5 mg/ml stock), 250-500 ng BSA, then dH2O up to 25 祃 volume. Incubate at ap......阅读全文
Streamlined－DNA－Extraction－Protocol
This method is derived from a procedure developed by Toby Bradshaw and the Poplar Molecular Genetics Cooperative. We have tested the procedure wit
RNA－analysis－on－nondenaturing－agarose－gel－electrophoresis
实验概要RNA analysis on non-denaturing agarose gel electrophoresis实验步骤1. The following gel electrophoresis conditions are recommended:- use 1X TAE buffer
RNA－analysis－on－nondenaturing－agarose－gel－electrophoresis
1. The following gel electrophoresis conditions are recommended:- use 1X TAE buffer instead of 1X TBE- use agarose gel in the concentration of 1.1%-1.
A－safe－method－of－extracting－DNA－from－Coccidioides－immitis
Human-pathogenic fungi such as Coccidioides immitis and Histoplasma capsulatum must be handled in Biosafety level 3 containment facilities which make
Restriction－Digest
Materials:Restriction enzymes of choice, such as BamH1 and EcoRIRestriction enzyme reaction buffer, such as MULTI-CORE (TM) (Promega)70 % Ethanol100 %
Restriction－digestion
Restriction digestionRestriction enzyme digestions are performed by incubating double-stranded DNA molecules with an appropriate amount of restriction
ADPRibosylation－Factor
ADP-ribosylation factors (ARFs) are 20-kDa guanine nucleotide-binding proteins, members of the Ras GTPase superfamily that were initially recognized a
Roche公司的RNase－Protection－Assay－(RPA)－protocol
Roche公司的RNase Protection Assay (RPA) Using DIG-Labeled RNA Probes下载网址：http://www.roche-applied-science.com/PROD_INF/BIOCHEMI/no1_03/PDF/p22_23.pdf还有一份
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
Complete－PCR－Guide
In the polymerase chain reaction (PCR), a thermostable DNA polymerase amplifies DNA that is flanked by known sequences. The known sequences correspond
DNA转化实验指导3
6. Simultaneous digestion of the pUC vector with both enzymes in the presence of 3 units of Shrimp Alkaline Phosphatase (Amersham BioSciences) in
Restriction－Enzyme－Buffer
Restriction Enzyme Buffer Most enzymes can use REact buffers; however, some are made up separately. Use fresh Milli-Q water, siliconized or sterile gl
The－Restriction－Enzyme－Database
REBASE The Restriction Enzyme Database The Restriction Enzyme data BASE A collection of information about restriction enzymes and related proteins. It
Lambda－DNA－Preparation
Lambda DNA PreparationThis is a plate method that gives very good yield for cloning. I have combined the Promega and Maniatis protocols.Solutions T-TY
EGel®－CloneWell－Agarose－Gels
实验概要Instructions are provided below for using the E-Gel®CloneWell pre-cast agarose gels with the E-Gel® iBase™ Power System. For detailed instructio
DNA－Purification－from－Agarose－Gels
1. Separate DNA fragments in an agarose gel cast with 0.5 mg/mL Ethidium bromide. Locate bands with a hand-held long-wave UV lamp.2. Slice the gel wit
Genomic－DNA－Extraction－－PureLink™
实验概要The PureLink™ Genomic DNA Purification Kit allows rapid and efficient purification of genomic DNA. The kit is designed to efficiently isolate g
Detection－of－apoptotic－process－in－situ－using－immunocytochemical
1. INTRODUCTION Apoptosis was observed from invertebrates to lower and higher verterbrates, and intervenes both in physiological and in pathological
Extraction－of－DNA－using－DNAzol®－Reagent
实验概要DNAzol® Reagent (Genomic DNA Isolation Reagent) is a complete and ready-to-use reagent for the isolation of genomic DNA from solid and liquid sa
Yeast－Media
YEPD (non-selection)-1% yeast extract-2% peptone-1.5% agar (if needed for plates)After autoclaving add glucose to 2% by adding 100 ml of 20% solution
细胞培养常规操作
常规操作(主要内容如下)· Aseptic Technique· Culture Vessels· Cell Counting· Primary Culture· Maintenance of Cell Line ·
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 (
巨噬细胞和单核白细胞
· Lymphocyte Transformation (Donis-Keller lab)Lymphocytes are transformed to establish cell lines. Mononuclear cells (lymphocytes) from antico
Method:－Preparation－of－Lymphocyte－Cell－Pellet－for－Storage
Method: Preparation of Lymphocyte Cell Pellet for StorageJune 10, 1990Rosalie VeilePurpose:Following propagation to 1 X 108 cells, lymphoblastoid cell
Methylation－Specific－PCR
Methylation Specific PCRProtocol written by James Herman*Methylation Specific PCR (MSP) is a simple rapid and inexpensive method to determine the meth
DNA分子的限制性内切酶（Restriction－enzyme－,－endonuclease）消...
限制性内切酶可特异地结合于一段被称为限制酶识别序列的DNA 序列位点上并在此切割双链DNA 。绝大多数限制性内切酶识别长度为4、5或6个核苷酸且呈二重对称的特异序列，切割位点相对于二重对称轴的位置因酶而异。一些酶恰在对称轴处同时切割 DNA 双链而产生带平端的DNA 片段，另一些酶则在对称轴
Ligation－Optimization
The following protocol can be used to optimize ligation conditions for difficult to clone (e.g. very large) fragments. The principle is to independent
High－Efficiency－Transformation
Day 0Make sure you have the necessary solutions (instructions for how to make them can be found here):Single-stranded carrier DNAPEG 3350 50% w/vol1.0
Modified－Yeast－Transformation
Inoculate cells from an overnight culture into 50 ml YEPD and incubate at 30°C with shaking. Typically, add 0.1 to 0.2 ml saturated culture in the eve
Fast－Yeast－Transformation
Protocol: Fast yeast transformationAdd 50 µl carrier DNA to a 1.5 ml tube.scrap cells from plate and add to the carrier DNA.Add in the following order