HowdoyousynthesizeyourdsRNA
We routinely produce dsRNA by in vitro transcription of a PCR generated DNA template containing the T7 promoter sequence on both ends (I. Primer Designed dsRNA). It is also possible to produce dsRNA using PCR generated DNA templates containing either the T7 & SP6 or the T7 & T3 promoters on either end (II. dsRNA Fom Clones). This method is less efficient, especially when working on a large scale.All work shou......阅读全文
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
How－do－I－decontaminate－my－tissue－culture－(Invitrogen)
When an irreplaceable culture becomes contaminated, researchers may attempt to eliminate or control the contamination. First, determine if the contami
Creation－and－Use－of－Infectious－Virus－Vector
Creation and use of your infectious vector:Plate 5 x 105 293T cells in 6 cm2 dishes containing 5 ml of media. (This can be scaled up if desired).The f
Degenerate－PCR,－a－short－guide.
What is degenerate PCR? Degenerate PCR is in most respect identical to ordinary PCR, but with one major difference. Instead of using specific PCR pr
CREATION－AND－USE－OF－YOUR－INFECTIOUS－VECTOR
实验概要CREATION AND USE OF YOUR INFECTIOUS VECTOR实验步骤Day 1 1. Plate 5 x 105 293T cells in 6 cm2 dishes containing 5 mL of media. (This can be scal
Protein－Expression－and－Purification－Protocol
Step 1: Transform appropriate DNA plasmid into BL21(DE3) E. coli cells. These cells must be competent. (Protocol for how to make competent cells.)a) T
Gel－Electrophoresis－of－DNA
What is Electrophoresis?Electrophoresis is a technique used in the laboratory that results in the separation of charged molecules. In this CyberLab we
Belcher/Knight:－Electrocompetent－Cells
Electrocompetent CellsFrom Danijela Dukovski at Harvard Medical School. This protocol works well. --Julie NorvilleMaterialsDI water10% GlycerolSpecial
Bench－Top－Radioactive－Work－Protocol
Log in radioactive material received and deduct amount of radioactive material used in each experiment in Radioisotope Log Book.Clear your bench top w
Degenerate－PCR
Degenerate PCR is in most respects identical to ordinary PCR, but with one major difference. Instead of using specific PCR primers with a given sequen
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
Basic－Method－for－Indirect－Immunofluorescence－Labeling
Basic Method for Indirect Immunofluorescence LabelingBackgroundThis is the method for indirect immunofluorescence labeling; that is, the antibodies do
sothing－about－Genome－walking
Can anyone recommend a good kit for genome walking? I would like to find out the promoter sequence of a known gene. Is genome walking the way to do it
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Do you rely on tank scales when filling, batching or tracking inventory? If so, accuracy limitations can diminish product quality, leading to mate
软件技术如何帮助开发更精确有效的体外诊断系统？
The use of in vitro tests often plays an important role in the diagnosis and treatment of disease. However, many of these tests rely on detecting
HOW－TO－USE－THE－COULTER－COUNTER－TO－COUNT－CELLS
1) Turn on the counter by pulling out the on/off button. You need to do this at least 10 min before use to obtain sufficient vacuum.Usually put 0.2 ml
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Phosphoamino acid analysis:Mark Kamps's method1. Label your protein with 32Pi. Then subject the protein to SDS polyacrylamide gel electrophoresis
PCR－Primers－For－Gene－Expression－Detection－or－Quantification
Why PrimerBank?PrimerBank is a public resource for PCR primers. These primers are designed for gene expression detection or quantification (real-time
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Good Pipetting PracticePipetting Techniques to Boost Your PerformanceImprove your data quality with Good Pipetting Practice™ (GPP™) – METTLER TOLE
annealing－temp－for－degenerated－primer－－PCR－problem
PCR primers should be free of significant complementarily at their 3'-termini as this favours formation of primer-dimer which reduce product yield
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
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
针对特定细胞选择更有效的实验步骤
GenJet Ver.II，LipoD293及PolyJet针对哺乳动物细胞的DNA体外转染试剂，均为您提供两种不同的转染步骤——一般步骤及高级步骤，这两种步骤分别针对不同的哺乳动物细胞。高级步骤主要针对难转的哺乳动物细胞，例如：MDCK，MDA-MB231，Caco-2等细胞。使用一般的转染步
FIXATION－and－DNA－Staining－for－Cell－Cycle－Analysis
BackgroundThis method of DNA staining utilizes ethanol to fix the cells and permeabilize the membrane, which allows the dye (Propidium Iodide) to ente
ProcartaPlex－免疫检测常见问题及解决方案
eBioscience公司Luminex液相蛋白芯片检测试剂——ProcartaPlex可以为您的生物学研究提供各种多因子检测方案，包括细胞因子、生长因子、趋化因子和其他蛋白。在选择试剂盒及操作时会遇到很多问题，现将常见问题及答复汇总如下，供您参考： 如果不小心将整个试剂盒保存在-20°C. 这个试
How－to－interrupt－scintillation
Peter Novick Lab, Department of Cell Biology Yale University School of Medicine HOW TO PERFORM AN INTERRUPT OF ANONGOING AUTOCOUNT1). This program int
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
如何选择麻醉机？
关于麻醉机如何选择型号，可以从以下几个方面寻找突破： 1. 实验过程中的动物种类以及实验动物（一只）的重量（比较重要）？ What species are you working with and what is the weight range (very important) of each
Tissue－Culture－Methods3
REFERENCES:R. Ian Freshney, Culture of Animal cells: A manual of basic techniques, Wiley-Liss, 1987.VI. TISSUE CULTURE PROCEDURESEach student should m
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