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N.B: Gloves should be worn at all times during preparation of in vitro RNA. All solutions should be RNase-free i.e. made with DEPC water if home-made or bought specifically to use for RNA work. You w ...

Feature The RNase A is free of DNase activity. It is not necessary to heat it before use. Description The Ribonuclease A (RNase A) is an endoribonuclease that specifically degrades single-stranded RNA ...

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！ A.Primer Designed dsRNA B.Template Selection C.PCR D.In vitro RNA Transcription E.dsRNA Purif ...

siRNA Design RNAi target selection rules : Targeted regions on the cDNA sequence of a targeted gene should be located 50-100 nt downstream of the start codon (ATG). Search for sequence motif AA(N19 ...

RNA i Glossary Dicer - Dicer is a member of the RNase III family of nucleases that specifically cleave double-stranded RNA s. Dicer processes long dsRNA into siRNA of 21-23 nt. Interferon - A small ...

1.NCBIshRNA资源库 http://cgap.nci.nih.gov/RNAi About RNAi on CGAP The NCBI is part of the consortium supporting the preparation of human and mouse libraries containing RNAi constructs that target cance ...

Polymerase III in vitro Transcription Steve Hahn For the following reactions use appropriate shielding and dispose of radioactive waste properly! A 20 microliter transcription reaction contains: 4.0 ...

Preparation of Cells 1.Prepare or collect between 2x107 cells for each mRNA prep (will yield about 10-20µg of mRNA). If PBMCs from a whole blood sample are to be used the sample should be prepar ...

pSico Oligo Design Click on the Mac OSX program pSicoOligomaker 1.5 to select and design oligos for pSico and plentilox3.7. To operate it simply paste your sequence in the "sequence" window ...

Run-on transcription monitors the regulation of transcription in isolated nuclei. A. Preparation of Nuclei - (do everything at 0℃ to 4℃ in 50 ml tubes) 1. Pellet between 30 and 300 million cells at 15 ...

T℃OS-M6 cells grow in 6-well plates 2ml media total T℃V1PD cells grow in 100mm plates. 1: Aspirate off the media from the cells and was the cell monolayers with 2x5ml (To 100mm dish) or 2x1ml (to 6-w ...

Add 0.1 ml DEPC to 100 ml of the solution to be treated and shake vigorously to bring the DEPC into solution. Let the solution incubate for 12 hours at 37℃. Autoclave for 15 minutes to remove any trac ...

3' R apid A mplification of c DNA E nds (RACE ) PCR This technique is used to obtain the 3'end of a cDNA it requires some sequence information internal to the mRNA under study. The sequence inf ...

1) Harvest 1x105 to 108 cells. Wash 1x w/PBS and freeze in liquid N2 and store @ -70℃ 2) Resuspend each pellet in 1.5ml lysis buffer (300μl 5x Lysis 75μl 200mM VR 1.125ml H2 O 3) Incubate on ic ...

Steve Hahn. last modified Sat Oct 17 1998 Mix the following in an 0.5 ml eppendorf tube: 10-20 micrograms total yeast RNA 10 microliters hybridization mix H2O to bring the final volume to 25 microlite ...

1 ml linearized DNA template pBS-SK- ERD (X-E) (0.1-0.5 mg) 7.8 ml DDW 5 ml 5X transcription buffer ( stratagene ) 1 ml 750 mM DTT 1 ml Rnasin (RNase inhibitor; 15U/ml) 1 ml 10 mM ATP (pH 7) 1 ml 10 m ...

Ambion and Applied Biosystems have joined forces to provide a complete convenient solution for performing gene silencing experiments and validating the results by real-time RT-PCR. Ambion's Silencer&t ...

READ THROUGH ALL CAUTIONS BEFORE TRYING THIS EXPERIMENT Materials •Rat liver (fasted rat) •Liquid nitrogen •p-Amino-salicic acid •Phenol mixture •Homogenizer or blender6 &bull ...

RNase Protection Assay (RPA) BD Biosciences公司， PDF文件 http://www.bdbiosciences.ca/canada/downloads/protocols/RPA.pdf ...

The use of small interfering RNAs (siRNAs) to induce targeted gene silencing in mammalian cells offers researchers a powerful tool for analyzing gene function. Ideally one would like to work with indi ...