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ChIP-Chip E. coli: Difference between revisions
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==Materials== | ==Materials== | ||
*Ultralink protein A/G beads (catalog number 53132, [http://www.piercenet.com/ Pierce]) | *2 ml Ultralink protein A/G beads (catalog number 53132, [http://www.piercenet.com/ Pierce]) | ||
*Specific antibody for example RNA polymerase β subunit from [http://www.neoclone.com/productcatalog/index.php Neoclone, Madison] | *Specific antibody for example RNA polymerase β subunit from [http://www.neoclone.com/productcatalog/index.php Neoclone, Madison] | ||
===Reagents=== | ===Reagents=== | ||
Material for about 50 experiments: | |||
75 ml Formaldehyd (37%) | |||
500 ml 2.5 M Glycine | |||
1.5 l [[TBS]] (400 ml 5xTBS) | |||
[[ChIP-Chip E. coli/Lysis Buffer |50 ml Lysis Buffer]] | |||
[[ChIP-Chip E. coli/IP-Buffer |IP-Buffer (Immunoprecipitation Buffer)(500ml)]] | |||
2 ml [[PMSF]] | |||
30 ml I-Buffer + 500 mM NaCl | |||
30 ml Wash-Buffer: | |||
10mM Tris pH 8.0 | |||
250 mM LiCl | |||
1 mM EDTA | |||
0.5% Nonidet-P40 | |||
0.5% Sodium deoxycholate | |||
50 ml TE | |||
10mM Tris pH7.5 | |||
1mM EDTA | |||
10 ml Elution-Buffer: | |||
50 mM Tris (pH 7.5) | |||
10 mM EDTA | |||
1% SDS | |||
20 mg/ml Pronase in TBS (store up to 1 year at -20 C) | |||
===Equipment=== | ===Equipment=== | ||
Any equipment used to perform the protocol (link to a method for using them). | Any equipment used to perform the protocol (link to a method for using them). | ||
