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
SYBR-Green-Quantitative-PCR-Protocol
SummaryQuantitative PCR is a method used to detect relative or absolute gene expression level. All qPCR involves the use of fluorescence to detect the
禽流感病毒RNA提取protocol
1,用异硫氰酸胍提取1.取200ul样品数+阴性对照+阳性对照个1.5ml灭菌eppendorf管2.加600ul异硫氰酸胍,然后加入对照和样品,再加200ul氯仿,颠倒混匀3.13000rpm离心15min4.在第3步离心快结束时,另取同样多eppendorf管,加入400ul -20度预冷的异丙
病毒RNA提取实验方法(protocol)
1,用异硫氰酸胍提取提禽流感病毒的详细步骤,可参考(我提过N次做定量PCR都没问题):1.取200ul样品数+阴性对照+阳性对照个1.5ml灭菌eppendorf管2.加600ul异硫氰酸胍,然后加入对照和样品,再加200ul氯仿,颠倒混匀3.13000rpm离心15min4.在第3步离心快结束时,
酵母转化的几种方法
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 m
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
Miniprep/Qiagen-kit
MaterialsFor purifying plasmid DNA from Escherichia coli cells, the Qiagen Spin Miniprep Kit produces quite reliable results.Do not autoclave solution
线虫总RNA提取及RTPCR实验方法
C. elegans RNA Isolation and RT-PCRReagents Needed:M9 (common stock)Trizol (stored at 4ºC)chloroform2-propanol70% EtOHRNase-free H2OiScript cDNA Synth
E.Z.N.A.™-Mollusc/Arthropod-RNA-Protocol
实验概要The E.Z.N.A.® Mollusc RNA Kit is designed for efficient recovery of total RNA greater than 200 nt from molluscs, arthropods, roundworms, flatw
细胞遗传学——原位杂交(ISH)
In Situ Hybridization· In Situ Hybridization (jsmith1@po-box.mcgill.ca)In situ hybridization, as the name suggests, is a method of localizing,
ORNL-MICROARRAY-HYBRIDIZATION-PROTOCOLS
Direct labeling of total RNA with Cy3 and Cy5:A. MATERIALSRNeasy® Mini Kit (Qiagen; Cat # 74106) SuperScript II RT (200U/µL) (Life Technologies; Cat #
Purification-of-Genomic-DNA-Using-PureLink™-Silica-Columns
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 ge
Cloning-of-small-RNAs-with-5’-phosphate-and-3’-OH-ends3
Load your precipitated PCR samples into 2 consecutive lanes so as not to overload the lanes. For each different sample, I would run a separate ladder
DNA-and-RNA-EXTRACTIONS
A protocol / method / schedule /procedure for extraction / isolation of both DNA and RNA from the same material typically plant leaf / leaves(See also
Bioanalyzer
Protocol for Bioanalyzer RNA nano chippreparation of material12 samples per chipquantitative range 25–500 ng/μlquantitation accuracy 20%CV (for ladder
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
果蝇RNAi的实验中双链短RNA的合成(dsRNA)方法
实验概要We routinely produce dsRNA by in vitro transcription of a PCR generated DNA template containing the T7 promoter sequence on both ends (I. Prim
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
果蝇RNAi的实验中双链短RNA的合成(dsRNA)方法
本文来自于哈佛大学医学院果蝇RNAi筛选中心的经典实验方法,专门用于果蝇RNAi实验方法。感谢哈佛大学医学院果蝇RNAi筛选中心的支持!Primer Designed dsRNATemplate SelectionPCRIn vitro RNA TranscriptiondsRNA Purifica
BIOG-RNA-Swab-Kit(拭子RNA提取试剂盒)使用说明
试剂盒组成 组分 50次 100次 吸附柱和收集管 各50个 各100个
E.coli-Total-RNA-Labeling-Protocol-for-Spotted-Microarray
Note:Start with 20 ug of total RNA for each labeling reaction.All solutions that can be filtered should be filtered.Cy dyes are light sensitive and sh
Largescale-Immunocytology
This protocol describes our method for preparing cells for immunofluorescence, in which all incubations and washes are performed in microtiter dishes.
Isolation-of-Total-RNA-from-Animal-Cells-use-RNeasy-Mini-Kit
实验概要Extract the total RNA from animal cells by using RNeasy Mini Kit (QIAGEN No.74104) 主要试剂SDS based extraction solution实验步骤1. Harvest cells.1) Try
Protocol-for-dsRNA-Synthesis
实验概要 We routinely produce dsRNA by in vitro transcription of a PCR generated DNA template containing the T7 promoter sequence on both ends
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
How-do-you-synthesize-your-dsRNA
We routinely produce dsRNA by in vitro transcription of a PCR generated DNA template containing the T7 promoter sequence on both ends (I. Primer Desig
Yeast-Media
YEPD (non-selection)-1% yeast extract-2% peptone-1.5% agar (if needed for plates)After autoclaving add glucose to 2% by adding 100 ml of 20% solution
Plasmid-isolation-from-yeast
Pick colonies into 0.5ml of SD-Leu (or other appropriate SD medium)Vortex for 1minLeave to grow O/N for 18-24h at 30°C, 230-250rpm (best in 5ml bijou)
DNA-Immunoprecipitation-for-the-Determination-of-DNABinding-Specificity
Andrea J. Gossett and Jason D. Lieb1Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA1Corresponding autho
酵母转化
· Yeast Transformation (Gietz Lab)LiAc/SS-DNA/PEG Transformation· Yeast Transformation (Breeden Lab)LiAc method· Large-Scale Y
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,