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PCR Transformation
日期：2006-08-17 06:28:24 点击：475 好评：0PCR Transformation Grow cells to an OD600 0.7-2.5 (25ml culture) Spin cells 4500 rpm for 2 min Resuspend in 50 ml 0.1M LiOAc Repeat step 2 and 3 Resuspend in 0.1M LiOAc (culture volume x OD/100) Add 1/9 volume of 10 mg/ml ssDNA that has be...
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Isolation of total cytoplasmic RNA
日期：2005-07-18 00:00:00 点击：577 好评：0Description Isolation of total cytoplasmic RNA using LiCl-Urea extraction and PK digestion – Auffray and Rougeon modified method RNA extraction suitable for RNases rich tissues Procedure Sacrifice animals by cervical dislocation. Dissect...
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RNAi in HeLa cells
日期：2005-07-18 00:00:00 点击：627 好评：0Sunday, February 08, 2004 Description This is a popular procedure, in fact, it could be the most popular mammalian genetic technique out there right now. The only issue is that most labs seem to do it differently with variable success. Her...
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Ribonuclease Protection Assay
日期：2005-07-18 00:00:00 点击：518 好评：0Wednesday, November 19, 2003 Description Most RNase Protection protocols require an overnight hyb with numerous subsequent clean-up steps. This method requires a maximum hyb of four hours, and the clean-up steps are the barest minimum, yet...
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RNA synthesis from plasmid-borne sequences
日期：2005-07-18 00:00:00 点击：458 好评：0Tuesday, November 18, 2003 Description In vitro RNA synthesis from plasmid-borne sequences under the control of T phage promoters Procedure 1) Linearize the plasmid at, or near, the terminus of the insert cDNA with a suitable restriction e...
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Yeast Chromatin Immunoprecipitation
日期：2005-07-18 00:00:00 点击：505 好评：0Tuesday, November 18, 2003 Description Yeast Chromatin Immunoprecipitation Procedure Day 1 1. Start 5 ml o/n cultures of strains to be tested Day 2 1. Inoculate 50 ml of the desired media with a volume of a saturated o/n culture and grow o...
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Single Stranded dut ung DNA Prep
日期：2005-07-18 00:00:00 点击：423 好评：0Tuesday, November 18, 2003 Description Single Stranded dut ung DNA Prep Procedure Transform the plasmid to be mutagenized into strain CJ236 (dut- ung-). Patch several transformants to YT amp +chloramphenicol (30 micrograms/ml; make fresh c...
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Primer Extension analysis of total yeast RNA
日期：2005-07-18 00:00:00 点击：368 好评：0Tuesday, November 18, 2003 Description Primer Extension analysis of total yeast RNA Procedure 1. Mix the following in a 1.5 ml eppendorf tube: a. 20-80 micrograms yeast RNA dissolved in H20 (the amount will depend on the level of expressio...
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Seed RNA Extraction Protocol
日期：2005-07-18 00:00:00 点击：476 好评：0Monday, November 10, 2003 Description Seed RNA Extraction Protocol Procedure After harvesting, siliques should be immediately frozen in liquid N2. Samples are stored at -80C until the extraction Place all solutions, eppen etc on ice to coo...