HELPERPHAGEPREPARATION
HELPER PHAGE PREPARATION1. Grow an overnight of NM522 in NZCYM medium.2. Dilute overnight 1:100 and grow to an A600 = 0.3 (@2.5 x 108 cells/ml).3. Infect cells with VCS helper phage to a m.o.i. = 0.1 (phage:bacteria = 1:10). There should be a high-titer stock of VCS in the lab.e.g., 40ml NM522 (@1010 cells) + 10ml VCS (@109phage from a stock = 1011 phage/ml)4. Incubate/shake @ 37oC for 8h.5. Centrifuge 10', 15krp......阅读全文
Competent－Cell－Preparation
实验概要Competent cells are those that possess more easily altered cell walls that DNA can be passed through easily. These cells readily incorporate f
Preparation－of－Agar－plates
Prepare media and add 1.5 agar before autoclaving it (15g per liter). After autoclavation, cool the media in a 55 degree waterbath. Do not allow the s
Metaphase－chromosome－preparation
Materials: RPMI 1640 medium fetal calf serum (FCS), 20% Colcemid (e.g. Boehringer Mannheim cell biology reagents, Best.-Nr. 295892) cell cuture flask
DGK－Membrane－Preparation
Reagents:Bacterial strainE. coli N4830/pJW10LB amp media50 µg/ml ampicillinHigh salt bufferfor 1 L50 mM KH2PO4 6.8 g150 mM KCl 11.18 g50 mM sodium pyr
Preparation－of－Mouse－Neutrophils
实验步骤Mice：8-16 weeks old malePrewarm buffer to room temperatureBuffer A : Ca2 /Mg2 ‐free Hank’s buffered saline solution [HBSS; Invitrogen, Grand Isla
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.
PREPARATION－OF－MICROINJECTION－PIPETTES
INJECTION AND HOLDING PIPETTESThe glass capillary tubing used should be thin walled, borosilicate glass without a fibre.e.g. Clark Electromedical Inst
CELL－MEMBRANE－PREPARATION
I. Solutions: A. Ca and Mg free Phosphate Buffered Saline (PBS) solution, buffered with 0.02M Hepes. pH=7.4 B. Ca and Mg free PBS, buffered with
噬菌体的扩增和定量实验
Part 1杂交瘤、单细胞克隆和噬菌体展示技术是抗体发现的三种主流技术，其中噬菌体展示技术作为诺奖级别的技术，在生物创新医药研发过程中有着十分重要的作用，极大的加快了抗体类药物研发的进程。噬菌体展示技术不仅在新的抗体和多肽的发现及优化过程中有着核心的作用，还能和传统的动物免疫技术相结合，与纳米抗体、
如何优雅的完成噬菌体的扩增和定量实验
Part 1杂交瘤、单细胞克隆和噬菌体展示技术是抗体发现的三种主流技术，其中噬菌体展示技术作为诺奖级别的技术，在生物创新医药研发过程中有着十分重要的作用，极大的加快了抗体类药物研发的进程。噬菌体展示技术不仅在新的抗体和多肽的发现及优化过程中有着核心的作用，还能和传统的动物免疫技术相结合，与纳米抗体、
Sample－preparation－(analytical－gels)
Sample preparation and solubilization are crucial factors for the overall performance of the 2-D PAGE technique. Protein complexes and aggregates shou
Easy－YAC－Preparation－Method
YAC TRANSFORMATION OF C. ELEGANS USING TOTAL YEAST GENOMIC DNA[This method is described in The Worm Breeder's Gazette (1997) A. Davies and J. Shaw
Preparation－of－Sonicated－Human－DNA
Purpose:To break up high molecular weight human placental DNA into fragment sizes of 500 bp or less which can be used as competitor DNA in Southern an
Protocols－for－LCM－preparation－and－analysis
Protocols for LCM preparation and analysis I. Preparation, LCM and RNA/DNA extraction of Frozen Tissue SectionsA. EmbeddingB. CuttingC. StainingII. Pr
Preparation－of－tubulin2
DAY 2: Cycling preparation of MT protein.Keep the brains in an evacuated plastic ziplock bag buried in ice from the time of slaughter during transport
Tissue－preparation－protocol－for－ChIP
实验概要This protocol describes how chromatin is prepared from tissue, which can subsequently be used for chromatin immunoprecipitation (ChIP). It is re
Midiprep－preparation－of－Plasmid－DNA
实验概要The PureLink™ HiPure Plasmid DNA Midiprep Kit allows purification of 100–350 μg of high-quality plasmid DNA from 15–25 mL overnight E. coli cul
Large－Scale－Tubulin－Preparation
Tubulin is purified from bovine/porcine brain by two cycles of polymerization/depolymerization followed by removal of copurifying proteins on a phosph
Preparation－and－Staining－of－Paraffin－Sections
I. Fixation and Processing of Tissue for Paraffin SectionsA. Fixation of Tissues in 10% Neutral Buffered FormalinSacrifice animal by prescribed and ap
Preparation－of－nucleic－acid－probes
Preparation of nucleic acid probesIn standard nucleic acid hybridization assays the probe is labeled in some way. Nucleic acid probes may be made as s
Blood－Smear:－Preparation－and－Staining
Blood Smear: Preparation and StainingReference:Davidson, I. and Henry J., Clinical Diagnosis by Laboratory Methods, I. Davidsohn and J. Henry, eds., W
Maxiprep－preparation－of－Plasmid－DNA
实验概要The PureLink™ HiPure Plasmid DNA Maxiprep Kit allows purification of 500–850 μg of high-quality plasmid DNA from 100–200 mL overnight E. coli cu
Preparation－of－Lactobacillus－Competent－Cells
OverviewInstructions on how to prepare Lactobacillus plantarum competent cells before electrotransformation.MaterialsMRS mediaCulture of L. plantarum
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
Standard－Operating－Procedures－for－T1Phage－Testing－Assay
I. Introduction:This assay uses a lawn of phage-susceptible E. coli (DH10B) embedded in a layer of agarose. This top agarose lays on a bed of standard
Preparation－and－Staining－of－Frozen－Tissue－Sections
I. Preparation of Frozen Sections for SectioningMaterials neeed:2-methylbutane (isopentane)Liquid NitrogenDry icePeel-Away?/sup> base moldsFrozen tiss
Preparation－of－Broth－and－Plates,－etc.
Recipes: 1) LB BrothMake 16 gm of LB Broth Base (Gibco #M27800C) up to 800 ml in ddH2O. Swirl to dissolve, then add 110 µl of 10 N NaOH. Autoclave. 2
Large－Scale－Tubulin－Preparation——2
III. Pouring a 1L Phosphocellulose (PC) ColumnResin: Whatman P11 Cellulose Phosphate -- fibrous cation exchanger(1 gram of PC swells to about 4 ml pac
Preparation－of－Rat－Liver－Cell－Cytosol
These protocols should yield enough cytosol and organelles for 1-200 MT/Organelle motility assays.Solutions and Reagents Freshly removed or flash fro
Preparation－of－Agarose－Gels－for－DNA－separations
Weigh out the desired amount of agarose and place in an Erlenmeyer flask with a measured amount of electrophoresis buffer, e.g. for an 0.8% gel, add 0