E.Z.N.A.TMYeastRNAKitSpinProtocol
实验概要The E.Z.N.A.® Yeast RNA Kit allows convenient isolation of high-quality total RNA from a wide variety of yeast species. Up to 2 x 107 log-phase cultured yeast cells can be processed. The system combines the reversible nucleic acid-binding properties of HiBind® matrix with the speed and versatility of spin column technology to yield approximately 30 ug of RNA, with an A260/A280 ratio of 1.7-1.9. Purified RNA......阅读全文
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
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
Yeast－Nuclei－Isolation
This method gives yeast nuclei which look nearly purified microscopically. Nuclei isolated in this way do not give active transcription extracts when
体液样本核酸提取纯化
无细胞体液包括：血浆或血清，脑脊液 ，尿液，其他无细胞体液，细胞培养上清液。QIAamp Viral RNA Mini Kit —— 从无细胞体液中纯化病毒 RNA1)快速纯化高品质病毒 RNA（见图 1）2)无需有机提取或乙醇沉淀3)重复性好，产量高4)完全去除污染物和抑制剂图1 扩增血浆中的RN
RNA－FISH－on－cultured－cells－in－interphase2
Post-labeling DNA processing and purificationQiagen PCR clean up to get rid of unused oligonucleotides;Add 20µl cot1DNA, 10µl ssDNA to compete for rep
DNA标记
DNA标记（主要内容如下） DNA Labeling by Nick Translation Random Primed Labeling End-Labeling Purification of Labeled DNA Non-isotopic Labeling OthersDNA L
Extraction－of－RNA－from－Fibrous－tissues
实验概要E.Z.N.A.™ MicroElute® Total RNA Kit provides a rapid and easy method for the isolation of up to 50 ug of total RNA from small amount of cultured
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
Large－Scale－Plasmid－Preps:－Qiagen/Cesium－Method
Most plasmids can be adequately prepped by kits containing DNA binding columns. These columns do not do a great job of separating plasmid DNA from con
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
CDNA文库
CDNA文库（主要内容如下）· Construction of cDNA Library· Construction of Genome DNA Library· Library Screening OthersConstruction of cD
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
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
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
Decontamination－of－cells－from－the－yeast
I Destroy yeast1. Aspirate medium and wash cell in PBS.2. Incubate cells at 37oC for 5 min in non-diluted antibiotic-antimycotic.3. In
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
Isolation－of－microRNA－(miRNA)
实验概要 This protocol utilizes the powerful guanidine isothiocyanate–phenol:chloroform extraction method which allows the rapid isolation of t
RNA提取实验方法流程
总RNA（total RNA）和信使RNA(mRNA)的抽提（自己的总结）1、实验目的提取人体细胞的总RNA和mRNA。 2、本实验所需试剂1）、CNE-2细胞及培养细胞的一系列条件，2）、PBS缓冲液， TRIZOL试剂，3）、DEPC处理过的水、大、中、小Tip及1.5 ml EP管,4）、新的
RNA提取实验方法
总RNA（total RNA）和信使RNA(mRNA)的抽提（自己的总结）1、实验目的提取人体细胞的总RNA和mRNA。 2、本实验所需试剂1）、CNE-2细胞及培养细胞的一系列条件，2）、PBS缓冲液， TRIZOL试剂，3）、DEPC处理过的水、大、中、小Tip及1.5 ml EP管,4）、新的
免疫荧光
Immunofluorescence Technique (Spector Lab)protocol for immunofluorescence on cells Immunofluorescence Protocol (Walter Steffen)Methanol fixationForma
mRNA－Amplification－with－T7－RNA－Polymerase
MaterialsMessageAmp II aRNA Amplication Kit (Cat #1751, Ambion)100% EthanolRNA samples (e.g. 1 µg RNA per sample)ProcedureReverse transcriptionVerify
RNA提取实验方法流程
1、实验目的提取人体细胞的总RNA和mRNA。 2、本实验所需试剂 1）、 CNE-2细胞及培养细胞的一系列条件， 2）、PBS缓冲液， TRIZOL试剂， 3）、DEPC处理过的水、大、中、小Tip及1.5 ml EP管, 4）、新的氯仿、异丙醇和乙醇， 5）、mRNA分离
RNA－FISH－on－cultured－cells－in－interphase
IntroductionFluorescence in situ hybridization (FISH) has become a widely used method in genome and molecular genetic studies. The technique is highly
HP－Tissue－DNA－Maxi－Protocol
实验概要The E.Z.N.A.® HP Tissue DNA Maxi Kit is designed for efficient recovery of genomic DNA up to 60 kb in size from up to 2 grams of tissue samples.
从棉花组织中提取微量RNA的标准实验方法protocol
Prior to Extractions ・ Bake all necessary glassware, metal spatulas, mortars, and pestles overnight in a 200℃ oven after wrapping them in aluminum f
RNA－extraction－using－trizol/tri
RNA extraction with TRIzol (Invitrogen product name) or the equivalent TRI (Sigma-Aldrich product name) is a common method of total RNA extraction fro