ChIP-Chip E. coli: Difference between revisions

ChIP-Chip E. coli (view source)

297 bytes added , 20 April 2009

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-*500 ml '''IP-Buffer (Immunoprecipitation Buffer)''':
+{| border="3"
-**50 mM HEPES-KOH pH 7.5
+!Final concentration !! For 500ml
-**150 mM NaCl
+|+ '''IP-Buffer (Immunoprecipitation Buffer)'''
-**1 mM EDTA
+|-
-**1% Triton X 100
+|50 mM HEPES-KOH pH 7.5||
-**0.1 % Sodium deoxycholate
+|-
-**0.1 % SDS
+|150 mM NaCl||
+|-
+|1 mM EDTA||
+|-
+|1% Triton X 100||
+|-
+|0.1 % Sodium deoxycholate||
+|-
+|0.1 % SDS||
+|-
+|}
-*30 ml IP-Buffer + 500 mM NaCl
-*30 ml '''Wash-Buffer''':
+*IP-Buffer with 500 mM NaCl: add 2.8mL 5M NaCl to 40mL IP-buffer
-**10mM Tris pH 8.0
-**250 mM LiCl
-**1 mM EDTA
+{| border="3"
-**0.5% Nonidet-P40
+!Final concentration !! For 500ml
-**0.5% Sodium deoxycholate
+|+ '''Wash-Buffer'''
+|-
+|10mM Tris pH 8.0||
+|-
+|250 mM LiCl||
+|-
+|1 mM EDTA||
+|-
+|0.5% Nonidet-P40||
+|-
+|0.5% Sodium deoxycholate||
+|-
+|}
+{| border="3"
+!Final concentration !! For 500ml
+|+ '''Elution-Buffer'''
+|-
+|50 mM Tris (pH 7.5)||500μL 1M
+|-
+|10 mM EDTA||200μL 0.5M
+|-
+|1% SDS||1mL 10%
+|-
+|ddH<sub>2</sub>O||8.3mL
+|-
+|}
-*10 ml '''Elution-Buffer''':
-**50 mM Tris (pH 7.5)
-**10 mM EDTA
-**1% SDS
 '''For labeling the DNA:'''