ProtocolforCellFusion
Healthy Sp2/0 cells should be rapidly growing by this time. Sp 2/0 cells should be started about two weeks before the cell fusion. Every two days, they should be centrifuged at a 64.4 xg on the IEC clinical centrifuge (in 50 ml tubes) for two minutes. The media should be removed and the cells resuspended in fresh media. This ensures that only the healthiest cells will fall to the bottom of the tube. Centrifugation at......阅读全文
Fluorescence－Procedures－forthe－ActinandTubulin－Cytoskeleton－inFixed－Cells1
General StrategyWe typically work with tissue culture, primary mammalian cells, and cell extracts, but the protocols can be adapted to other systems,
Belcher/Knight:－Electrocompetent－Cells
Electrocompetent Cells From Danijela Dukovski at Harvard Medical School. This protocol works well. --Julie Norville Materials DI water 10% Glyce
Agrobacterium－growth－and－transformation
Growth and storage of Agrobacterium tumefaciensStrain GV3101: resistant to gentamycin and rifampicin so add 25-50 ug/ml Gentamycin, 10 ug/ ml rifampic
Dark－Field－Viewing
Dark Field ViewingPrincipleTo view a specimen in dark field, an opaque disc is placed underneath the condenser lens, so that only light that is scatte
Preparation－of－High－Titer－Adenovirus－in－P11－cells
adapted from Cell Biology, A Lab Manual, second edition, volume 1 -Grow up P11 cells in 15 cm plates to 70 – 80% confluence. -Infect cells with a MOI
DAPI－Counterstaining－Protocols
实验概要The blue-fluorescent DAPI nucleic acid stain preferentially stains dsDNA; it appears to associate with AT clusters in the minor groove. Binding
Miniprep/Kitfree－highthroughput－protocol
Background This protocol is adapted from "Molecular Cloning: A Laboratory Manual", Second Edition, Sambrook, Fritsch, and Maniatis. It is a quick, i
Realtime－PCR
实验概要The exponential amplification via reverse transcription polymerase chain reaction provides for a highly sensitive technique in which a very low
Expression－Library－Screening－(Procaryotic)－Using－APFusion－Proteins
Outline:Bacteriophage lambda is a linear double stranded DNA, approximately 50 kB in size. The two sticky ends help the phage to recircularize after e
Expression－L19－using－Pichia－pastoris
Pichia pastoris is a methylotropic yeast used to express high amounts of protein. Secreted expression is achieved with a cleavalable faktor. To yield
A－Method－for－Assaying－Deubiquitinating－Enzymes1
Abstract A general method for the assay of deubiquitinating enzymes was described in detail using 125I-labeled ubiquitin-fused αNH-MHISPPEPESE
Staining－Methods－for－cell－death
The simplest way: trypan blue. Dead cells stain blue Non-fixed cells: FDA(fluorescein diacetate)-green, alive cells; P.I. (propidium iodide)-r
Staining－Methods－for－cell
death Z. Xia 10/2/95 The simplest way: trypan blue. Dead cells stain blue Non-fixed cells: FDA(fluorescein diacetate)-green, alive cells;
Yeast－Cell－Cycle－by－Flow－Cytometry
ReagentsCold absolute ethanol.0.5 M Na citrate stock (filtered), 50mM diluted stock.10 mg/ml RNase A (Boil 10 mins, cool, filter and store at -20°C).4
自动化的微流控芯片系统在单细胞中检测MicroRNA的异质性3
结论· 我们在C1TM单细胞自动制备系统开发了一种简洁的实验方案，能以最少的手工操作，在不到24小时内，平行处理高达96个单细胞，对其miRNA表达谱进行分析。· C1 miRNA STA实验方案使用了Life Technologies为miRNA优化过的试剂。特别的，Megaplex™ RT及
The－Effects－of－Ultraviolet－Light－on－the－Fertilization
Jill K. Flemming, Franklin & Marshall College, Class of 2001IntroductionThe objective of this project is to observe the effects of UV radiation on bot
Collection－of－Peritoneal－Cells
Prepare a 10ml syringe fitted with a 26G short needle and filled with 5 to 7 ml of medium and 2 to 3 ml of air. Air is important. Prepare a Past
Collection－of－Peritoneal－Cells
Prepare a 10ml syringe fitted with a 26G short needle and filled with 5 to 7 ml of medium and 2 to 3 ml of air. Air is important. Prepare a Past
ELECTROPORATION－OF－ES－CELLS－AND－ISOLATION－OF－H/R－CLONES
Need 1.5-2 x 107 cells from a 2 day culture. 1. Cells are harvested as normal, washed x 1 in PBS then taken up at conc. of 1.2 x 10 7 cells/ml in co
ELISPOT－protocol
实验概要The procedure below is a general guideline procedure for ELISPOT. Abcam ELISPOT kits have been designed for detection of various cytokines and g
Cell－Cycles－－Introduction
The onion root tip and the whitefish blastula remain as the standard introduction to the study of mitosis. The onion has easily observable chromosomes
MINICHROMOSOME－MICROTUBULE－BINDING－ASSAY
Determine the OD600 and correlate the cell density from the chart. Set up four 100mL YPD cultures at the following densities: 0.7x105, 1x105, and 3
AMNIOTIC－FLUID－CULTURES－ON－COVERSLIPS
I. Purpose: Amniotic fluid may be used for prenatal diagnosis of aneuploidy or other structural abnormalities. II. Culture Procedure: A. Asepti
AMNIOTIC－FLUID－CULTURES－ON－COVERSLIPS
实验概要 AMNIOTIC FLUID CULTURES ON COVERSLIPS 主要试剂 Solutions: Colcemid working solution: 10 mcg/ml Colcemid in Hank's Balanced Salt Solution
E.Z.N.A.®－Plasmid－Maxi－Kit－Spin－Protocol
实验概要This Protocol is designed to isolate 500-1200 ug of high Copy-Number plasmids or 50-400 ug of low Copy-Number Plasmids from 200 ml overnight c
Purification－of－Lin,－ckit+,－Sca1+－bone－marrow－cells－for－Culture
MATERIALSMedia:Heat inactivated FBS (56°C x 30 min)PBS + 2% heat-inactivated FBSIMDM + 20% heat-inactivated FBSViral transduction: Iscove's + 20%
Subculturing－Adherent－Cells
实验概要The following protocol describes a general procedure for subculturing adherent mammalian cells in culture.主要试剂1. Complete growth medium, pre-warme
Procedure－for－Culturing－BG01V－Human－Embryonic－Stem－Cells
IntroductionHuman embryonic stem (hES) cells are pluripotent stem cells derived from pre-implantation embryos that can be maintained and expanded in a
Testing－for－Mycoplasma－by－Indirect－DNA－Stain－(Hoechst－33258－stain)
AimDNA staining methods such as Hoechst staining techniques are quick with results available within 24 hours, which compares favorably with 4 weeks fo
酵母GST蛋白纯化方法
GST Fusion Protein Purification from Yeast5 ml overnight culture of your favorite yeast in your favorite medium.Inoculate 50 ml and grow 30o C shaking