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Methylation PCR Primer Design
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日期：2008-03-23 03:37:59
点击：29 评论：0
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| OligoAnalyzer (IDT) Cool tool for oligo analysis. Features include analysis of hairpin, self-dimer, heter-dimer, and other basic parameters. http://www.idtdna.com/analyzer/Applications/OligoA Primer3 (Whitehead Institute/MIT Center for Genome Resear |
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DNA Methylation Analysis
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日期：2008-03-23 03:33:04
点击：15 评论：0
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| CpG Island Searcher (PA Jones Lab) The CpG island searcher screens for CpG islands which meet the criteria selected below in submitted DNA sequences. http://www.uscnorris.com/cpgislands/cpg.cgi CpG Plot (EBI) Detection of regions of genomic sequence |
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Bisulfite modification of DNA
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日期：2008-03-23 03:32:09
点击：21 评论：0
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| Day 1 Digest at least 5ug of DNA overnight with your favorite enzyme. Make sure that the enzyme does not cut within the region you plan to analyze. We have found that Bgl II works well for most of our analyses. I usually set up a 50 uL reaction. Run |
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Bisulfite Treatment of DNA
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日期：2008-03-23 03:30:26
点击：19 评论：0
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| Dilute DNA (up to 2 m g) into 50 m l with distilled H 2 O. Add 5.5 m l of 2M NaOH. Incubate at 37°C for 10 minutes (to create single stranded DNA). Add 30 m l of 10 mM hydroquinone (Sigma) to each tube, freshly prepared by adding 55 mg of hydroquin |
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DNA Methylation analysis
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日期：2008-03-23 03:26:32
点击：17 评论：0
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| Overview : DNA methylation is an epigenetic event that affects cell function by altering gene expression and refers to the covalent addition of a methyl group, catalyzed by DNA methyltransferase (DNMT), to the 5-carbon of cytosine in a CpG dinucleot |
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Immunoprecipitation of 35S Labeled Cells
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日期：2008-03-21 08:58:37
点击：20 评论：0
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| Cell Labeling Start with 1-5 x 10 6 cells that are in log phase growth For suspension cells, passage cells several hours prior to harvest for metabolic labeling. Cells that have recently been split may be in stationary phase and may not incorporate |
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Immunoprecipitation
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日期：2008-03-21 08:55:28
点击：10 评论：0
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| Outline Immunoprecipitation is a technique that permits the purification of specific proteins for which a antibody has been raised. This primary antibody is either already bound to agarose or can be bound to the protein A/agarose beads during the pr |
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IMMUNOPRECIPITATION
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日期：2008-03-21 08:53:44
点击：12 评论：0
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| MATERIALS: Running buffer: 2ul 1.25M Tris-HCl, pH 6.8 35ul distilled water 2.5ul 2-mercaptoethanol 12.5ul 10% SDS 10ul 80% glycerol 2ul bromphenol blue Make up running buffer fresh before use. TNE buffer: 10mM Tris-HCL, pH 8.0 10mM NaCl 0.5mM EDTA M |
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Immunoprecipitation from Eukaryotic Cell Extracts
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日期：2008-03-21 08:45:37
点击：23 评论：0
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| Grow cells to subconfluence. Aspirate media and wash with ice cold PBS twice and aspirate all of PBS. Lyse cells with ice cold PLC-lysis with protease +/- phosphatase inhibitors (see Reagents). Use 1.0ml per 10cm plate, 3.0ml per 15cm plate. Add PLC |
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Immunoprecipitation (IP) Protocol
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日期：2008-03-21 08:37:43
点击：30 评论：0
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| Overview : Immunoprecipitation (IP) is a method that uses the antigen-antibody reaction principle to identify a protein that reacts specifically with an antibody from mixture of proteins so that its quantity or physical characteristics can be examin |
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