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Documents 31 - 40 of 146 matches.
[10035more1.html]**
http://www.protocol-online.org/biology-forums/posts/10035more1.html
PCR AFTER SONICATION... - (OCT/04/2005 ) Pages: Previous 1 2  I didnt really change anything. It still workes eventhough the cells arent lysed b4 I sonicate. I just get more of a smear compaired to no ...
How can I decrease IgG background? - ChIP and Next Generation Sequencing**
http://www.protocol-online.org/biology-forums-2/posts/14067more1.html
... . Vigorously resuspend cell pellets in 300μl lysis buffer, incubate on ice for 10 minutes. 7. Sonicate for 15s*3 times at 45% output, with 1 minutes refractory period between sonications 8. Centrifugate ...
Expression in E.coli - Protein Expression and Purification**
http://www.protocol-online.org/biology-forums-2/posts/15618more1.html
... . -bhavana- This is how I do it: harvest 100 ml culture re-suspend pellet in 5ml lysis buffer, sonicate supernatant: soluble fraction pellet: re-suspend in 500 ul lysis buffer: insoluble fraction Use 1 ...
additional proteins in eluted protein-protein purification of GST tag fusion pro - Protein Expression and Purification**
http://www.protocol-online.org/biology-forums-2/posts/14742.html
... resuspended pellet 5mM dithiotheritol (DTT), 1mM EDTA and 0.25% sarkosyl added. The solution is sonicated 4 times (10secs of sonication with 10secs of cooling ice), speed 2 with Branson sonifier and centrifuged ...
SDS-PAGE - Protein and Proteomics**
http://www.protocol-online.org/biology-forums/posts/1330more1.html
... what protocol seems to give good clean lysate with SDS lysis buffer (re, specific steps such as sonication or heating etc). cheers! sean. -smath- I think the protein concentration of your sample may ...
Dual cross-linking thoughts - ChIP and Next Generation Sequencing**
http://www.protocol-online.org/biology-forums-2/posts/12734more1.html
... lowering the SDS concentration can improve the efficiency of the ChIP? create less bubbles during sonication step? I used Upstate ChIP kit. I tried DTBP before. It will enrich the DNA to which the protein ...
How can I decrease IgG background? - ChIP and Next Generation Sequencing**
http://www.protocol-online.org/biology-forums-2/posts/14067.html
... . Vigorously resuspend cell pellets in 300μl lysis buffer, incubate on ice for 10 minutes. 7. Sonicate for 15s*3 times at 45% output, with 1 minutes refractory period between sonications 8. Centrifugate ...
purity is so bad after elution from GST column - Protein and Proteomics**
http://www.protocol-online.org/biology-forums-2/posts/14634.html
... the proteins, though not all. Also, the DTT should be added before you lyse the cells (even before sonication). Another alternative is to tag both termini of the protein and do a double purification. ...
Spores separation from fermentation broth - General Lab Techniques**
http://www.protocol-online.org/biology-forums-2/posts/15827.html
... be greatly appreciated. -digestion- supposing you are working with fungal spores I would try (i) sonication or (ii) adding some mild detergent. For most fungi this should work (if the spores are not already ...
Fragment sizes after ChIP - ChIP and Next Generation Sequencing**
http://www.protocol-online.org/biology-forums-2/posts/14714.html
... have immunoprecipitated and I observe 2 smears, one at 100/250 pb and one at 1000/10000 pb. After sonication I have only a smear around 200bp (on agarose gel). So I wonder what is this second smear. Is ...

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