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Documents 51 - 60 of 143 matches.
negative control region issues - ChIP and Next Generation Sequencing**
http://www.protocol-online.org/biology-forums-2/posts/14181.html
... using WAY too much chromatin for each IP and might this be interferring with the IP? I have good sonication, probably an average of about 500 bp. -Dukey- It seems to me that increasing your amount of ...
Mock IgG versus No Antibody - Negative Control - ChIP and Next Generation Sequencing**
http://www.protocol-online.org/biology-forums-2/posts/12058.html
... thinking that perhaps I got high background because I didn't spin down my samples after sonication and before removing my chromatin for the IP. If it works out I'll update everyone in case someone ...
Laemmli Buffer Problems - SDS-PAGE and Western Blotting**
http://www.protocol-online.org/biology-forums-2/posts/14946.html
... what causes that, but assumed it may be DNA from the prep, or some other cellular debris. If I sonicate the pellets instead of boiling, I can get a higher concentration without the distortion. I've ...
homogenization devices - General Lab Techniques**
http://www.protocol-online.org/biology-forums-2/posts/10377.html
... , especially when using more buffer. How about a Mini-Potter or glass-on-glass tube. And should I sonicate afterwards. What does it change. By the way I use Triton x-100 and DTT in my buffer. thnx T -Tfal ...
Western blotting for proteins in vesicles? - Protein and Proteomics**
http://www.protocol-online.org/biology-forums-2/posts/12207.html
... the lysate because I didn't want to lose the membrane fraction. Before loading the lysates, I sonicated for 2 minutes. The problem is that the signal was still very weak, and I don't think I am ...
can a pellet be called "probe"? - General Lab Techniques**
http://www.protocol-online.org/biology-forums/posts/14703.html
... (100 mM) Lysozyme (final concentration 100 µg/ml) 2 µl 0.5 M EDTA pH 8.0 20 min at 4°C 3-4 min sonication bath spin 5 min at 14,000 rpm take supernatant and store at 4°C denature probe 5 min at 50 ...
Noise in ChIP-Chip data - DNA Methylation and Epigenetics**
http://www.protocol-online.org/biology-forums-2/posts/10955.html
... visual data. Let me precede all of this by saying I've done numerous controls and tests at the sonication, IP, and array steps, all of which appear to give me accurate data. However, when I visualize ...
TCA precipitation from High% sucrose solutions - Protein and Proteomics**
http://www.protocol-online.org/biology-forums-2/posts/14682.html
... in Sample buffer with 0.5M urea and 0.2M tris (8.8) (also BME, DTT, SDS) -RT O/N. -Full power sonication 5 minutes. -Boil 15 minutes. -Load, run...etc. In this process I seem to be losing about 50-75 ...
First ChIP - good start? - ChIP and Next Generation Sequencing**
http://www.protocol-online.org/biology-forums-2/posts/13098.html
... to the GLUT4 coding sequence as my negative control region since it should be silent in liver. Sonication looks nice with a good smear from around 700 - 250bp. Results (Ct) (Figure 1): GAPDH Input (10 ...
Whole cell lysis of fibroblasts - SDS-PAGE and Western Blotting**
http://www.protocol-online.org/biology-forums-2/posts/14983.html
... while for lysis to take place, or is resuspension enough. Usually what I do after resuspending is sonication in an ice bath and then centrifug. to collect the protein solution (supernatant). 2) My lysis ...

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