Decontaminationofcellsfromtheyeast
I Destroy yeast1. Aspirate medium and wash cell in PBS.2. Incubate cells at 37oC for 5 min in non-diluted antibiotic-antimycotic.3. Incubate cells at 37oC for 5 min in trypsin. Spin them down (100g, 5 min). DO NOT EXCEED 100g.II Secure cells from crosscontamination4. Slowly a......阅读全文
Glycolysis－Pathway
Glycolysis was one of the first metabolic pathways studied and is one of the best understood, in terms of the enzymes involved, their mechanisms of ac
Preparation－of－Yeast－DNA－Embedded－in－Agarose－Plugs
Preparation of Yeast DNA Embedded in Agarose PlugsAnja van Brabant(adapted from Iadonato, S. P., and A. Gnirke. 1996. RARE-cleavage analysis of YACs.
Invitro－Phagocytosis－Assay－of－Macrophages
IntroductionThe term phagocytosis itself describes its mean phage = engulfment; cytosis: cell process. In other words, phagocytosis is the cellular pr
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
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
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%
Preserving－yeast－cultures
Short term storageYeast cultures are stable for 1-2 weeks when refrigerated. Petri dishes should be sealed or in plastic bags.Medium term storageYeast
yeast:Assaying－mating
SetupYou have yeast strains that are deficient in mating (eg Ste12 knockouts) and would like to test whether transforming them with a plasmid that con
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
Yeast－DNA－Prep
Protocolgrow up yeast culture to appropriate density (near saturation)spin 1.5 mls of culture for 1 min in microfuge and aspirate off supernatantresus
Dropout－plates－for－yeast
Materials(Solutions are all available from the media room)200ml bottle of 2x SD200ml bottle of 4% agar -- make sure to sign it out40% glucoseCSM minus
人工转录因子的部件——人类锌指结构1
Human zinc fingers as building blocks in the construction of artificial transcription factorsKwang-Hee Bae1, 4, Young Do Kwon1, 2, 4, Hyun-Chul Shin1,
酵母遗传学技术
Genome-wide Gene Expression Analysis (Richard Young Research Group,Whitehead Institute for Biomedical Research)A genoe-wide gene expression analysis u
Nitrogendepedent－regulation－of－Rtg1－and－Rtg3－in－TOR－pathway
Many key signaling molecules are conserved from yeast to man. mTOR is a protein kinase involved in nutrient and growth factor signaling in humans that
Timing－of－Cycles
MaterialsMonolayer cultures grown in 75 mm culture flasks (Cells from Exercise 11.4 may be used, or cultures of tetrahymena, yeast, or algae may be us
蛋白质提取和纯化
蛋白质提取和纯化（主要内容如下）Protein Extraction Protein PurificationProtein PrecipitationColumn PreparatioinQ & A Posted in the Method ForumProtein ExtractionWhole
Largescale－Immunocytology
This protocol describes our method for preparing cells for immunofluorescence, in which all incubations and washes are performed in microtiter dishes.
酵母染色体沉淀分析方法
ABSTRACTThis protocol describes a method for the detection of proteins bound to specific regions of chromatin in yeast. There are many variations of t
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
DNA微序列技术
· Protocols for Making Drosophila Arrays (Stanford U.)Detailed protocol for making arrays including PCR Amplification of cDNAs for Printing,
Endy:Yeast－Colony－PCR
MethodUsing sterile pipette tips, transfer a 1 mm colony into 50 uL of 60 U/ml Zymolyase3 uL of 1 U/mL Zymolyase stock solution47 uL of waterIncubate
Blackburn:Yeast－Colony－PCR
OverviewThis is a quick and easy yeast colony PCR protocol that does not require zymolyase step.Updated Protocol: Blackburn Lab: Quick and Easy Yeast
Live－Cell－Imaging－of－Yeast
Live Cell Imaging of YeastDaniel R. Rines, Dominik Thomann, Jonas F. Dorn, Paul Goodwin and Peter K. SorgerINTRODUCTIONThe development of cloning vect
Yeast－Media,－Solutions－and－Stocks
Yeast Media:Note: Synthetic complete medium can be prepared by adding media supplements (see below).Medium using 6.7 g yeast nitrogen base without ami
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
Differential－Interference－Contrast
Differential Interference Contrast (Nomarski, DIC, Hoffman Modulation Contrast)PrincipleDifferential interference microscopy requires several optical
High－Molecular－Weight－Yeast－Liquid－DNA－Preparation
Purpose:To isolate intact, high molecular weight DNA from yeast cells for subcloning and rare cutting restriction enzyme analysis. One can expect a yi
线粒体荧光探针大全：TMRM，Mitotracker，JC1（3）
Mitochondrion-Selective Rhodamines and RosaminesRhodamine 123Rhodamine 123 (R302; FluoroPure Grade, R22420; Figure 12.22) is a cell-permeant, cationic
NUF2基因突变与药物因子介绍
该基因编码一种与酵母nuf2高度相似的蛋白质，nuf2是与着丝粒相关的保守蛋白复合物的组成部分。当着丝粒与纺锤体失去连接时，酵母nuf2在减数分裂前期从着丝粒中消失，并在染色体分离中起调节作用。该编码蛋白与有丝分裂hela细胞的着丝粒相关，提示该蛋白是酵母nuf2的功能同源物。已经描述了编码相同蛋白