DryTransfer干法蛋白轉膜
Trans-Bot SD Assembly1. Prepare the transfer buffer.2. Following electrophoresis, equilibrate the gels in transfer buffer. Equilibration facilitates the removal of electrophoresis buffer salts and detergents. The length of time required for equilibration is dependent on the gel thickness. For example, 15 min. for a 0.75 mm SDS-PAGE gel.3. Cut the membrane to the dimensions of the gel. Wet the ......閱讀全文
Isolation－of－genomic－DNA－from－bacteria
Note: This procedure does not work well with Gram + cocci.Transfer 1.5 mL overnight culture to a 1.5 mL microfuge tube, centrifuge for 30 sec, decant
Partial－Endonuclease－Digestion
Partial Endonuclease DigestionPrepare a 100 μl reaction mixture containing the DNA of interest in the appropriate 1X restriction enzyme buffer. Divide
CGH－Protocols－（一）
Metaphase chromosome preparationMaterials:?RPMI 1640 medium?fetal calf serum (FCS), 20%?Colcemid (e.g. Boehringer Mannheim cell biology reagents, Best
Yeast－Genomic－DNA－Prep
Grow 10ml YPD cultures o/n. Figure out cell density; inoculate 30 ml YPD and grow o/n so that cell density is approximately 2 x 108cells/ml the next m
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
Yeast－Lysates－for－Westerns
Cells are grown for 2-3 days as 1.5ml prep. under selection for the plasmid of interest. Spin cells down 2.6K for 5min.Resuspend in 1ml 0.25m NaOH/1%
愛丁堡集成Optistat－Dry到熒光光譜儀－測量溫度降至低于3K
分析測試百科網訊 愛丁堡儀器最近升級了現有的FLS980熒光光譜儀，使其可以在低于3 K到300 K的大的溫度范圍內進行測量，而不需要低溫液體如液氮或是更稀有的液氦。通過集成牛津儀器的新型光譜學恒溫器Optistat Dry，這成為了可能，Optistat Dry使用帶有氦氣閉合回路的Giffo
Rapid－DNA－Isolation－from－Phyllanthus－Amarus－and－Other－Plant－Tissues
Procedure Preheat Extraction Buffer at 60°C. Weigh 100 mg of fresh leaf tissue and grind it to powder in Liquid Nitrogen in a chilled mortar an
Western－Blotting－Protocols
back to topProtocolStandard vs. Rapid Immunodetection ProceduresThere are two types of protocols for immunodetection: Standard and rapid.Standard vs.
RLGS－protocol
A. Preparation of DNA SolutionIn the case of rice, for example This method may be appllicable for many grass species and some other plants.????????
RNA實驗方法3
Part III: HybridizationTurn heatblock on to 95C.For samples in water or ethanol, dry down appropriate amount of RNA, and include a tube with 1ul of tR
PCR－clean－up－using－3M－sodium－acetate－and－chilled－absolute－ethanol
For each 20 ul of PCR product, prepare the following mixture in 1.5 ml Tube.2 ul of 3M sodium acetate (NaOAc) pH 4.540 ul chilled absolute ethanol.Tra
Genomic－DNA－Extraction－－Phenol－|－Chloroform
實驗概要This section provides a general protocol for genomic DNA extraction using phenol and chloroform.主要試劑1.?????? Glycogen (20 μg/μL)2.?????? 7.5 M NH4
Phosphoproteins－pr...
實驗概要The following procedure provides a method of detection of phosphorylated proteins.實驗步驟1.?To a sample of protein solution containing 1-100 ng of
Transplantation－of－Pancreatic－Islets－Intothe－Kidney－Capsule－ofDiabetic－Mice
Preparation of Islets for Transplant (Tx)Under an inverted microscope, hand-pick islets using a P200 pipetman and straight pipet tip from the cultured
Fungal－Midi－DNA－Kit－Optional－protocol
實驗概要This protocol is designed for isolation of genomic DNA from fresh, frozen, or dried specimens from fungal samples contains higher phenolic mat
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
TRIzol－Prep
Procedure1.? Homogenize cells (10 million) or tissue (50-100 mg) in 1 mL?TRIzol Reagent?(e.g. scrape and pass through 30G needle, dounce homogenize an
細胞膜蛋白質提取方法
NRC Institute for Biological SciencesTriton X-114 extraction protocol (Hydrophobic protein preparation)Ressuspend cells in Solution A (dil 1/8) and ad
DNA－Extraction－from－Tissue
實驗概要DNA extraction from tissue.主要試劑Extraction buffer100 mM Tris-HCl (pH 8.0)?????100 mM EDTA (pH 8.0)?100 mM Na-Phosphate (pH 8.0)???1.5 M NaCl1% CTAB
Glycosphingolipid－analysis
1) Incubate cells with 1 μCi/ml of 3H-galactose for 72 hours.---> If treatment is for an extended period of time: treat in serum free media containing
Rat－Liver－Preparation
實驗概要The procedure presented below describes a method for preparing rat liver.主要試劑1.????? Aluminum Foil2.????? Liquid Nitrogen3.????? Dry Ice4.????? Ph
Plasmid－Miniprep
MaterialsSolution II: 0.2N NaOH/1% SDSSolution III: 3 M KOAc, pH4.8RNAseA (DNAse free) 10 μg/mLChloroform/Isoamyl alcohol (1/25 v/v)Isopropanol70 % et
Sphingomyelin－Quantitation－Postcholine－Labeling－of－HL60－Cells
Lipid Extraction1) Following the appropriate time of treatment, transfer 4.5 ml into each of two duplicate glass pyrex tubes and maintain on ice.2) Sp
James－Hardwick－CNBr－Cleavage－Procedure
1. Immunoprecipitate the protein and run it on a preparative gel. CNBr cleavage must be done with protein?transferred?to a nitrocellulose filter. Neit
Genomic－Southern－Blot
SolutionsProtocol:Digest 5-10 μg genomic DNA overnight with restriction enzyme of choice.Run digested gDNA on 0.8% TAE gel with marker (with no ethidi
酵母染色體沉淀分析方法
ABSTRACTThis protocol describes a method for the detection of proteins bound to specific regions of chromatin in yeast. There are many variations of t
RNEasy－Midi
MaterialsQiagen RNEasy Midi KitDounce homogenizers (for tissues)Syringes and needles (for cultured cells)70% EtOHSterile 15 mL conical tubesProcedure1
DNA－isolation－extraction
CTAB TECHNIQUE / Method / Schedule / Protocol FOR DNA ISOLATION / DNA EXTRACTION FROM PLANT LEAF / LEAVES SAMPLES (see also DNA RNA double isolation
Two－dimensional－peptide－mapping
This specfic protocol is the latest incarnation of peptide mapping procedures that have been developed here in the TVL/MBVL of the Salk Institute over