Yu:TEV Cleavage Buffer

(Difference between revisions)

Revision as of 15:33, 22 December 2009

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**Solution recipe for 1L TEV Cleavage Buffer.**
1. Measure 100 mL millipore H₂O into a 500mL beaker
2. Add 12.5 mL of 1M TRIS-HCl, pH 8.0	25 mM TRIS-HCl
3. Add 15 mL of 5M NaCl	150 mM NaCl
4. Add 500 μL of 0.5M EDTA	0.5 mM EDTA
5. Add 5 mL 10% NP-40	0.1% NP-40
5. Bring volume to 500L with millipore H₂O in a graduated cylinder	Volume to 1L
6. Immediately prior to use, complete with 1μL of 1M DTT per mL TEV CB	1 mM DTT

@@ Line 2: / Line 2: @@
 {| border="0" cellpadding="10" width="100%"
 |+'''Solution recipe for 1L TEV Cleavage Buffer.'''
-|width="66%" valign="top"|1. Measure 500 mL millipore H<sub>2</sub>O into a 1L beaker
+|width="66%" valign="top"|1. Measure 100 mL millipore H<sub>2</sub>O into a 500mL beaker
 |width="33%" valign="top"|
 |-
-|2. Add 50 mL of 1M TRIS-HCl, pH 8.0
+|2. Add 12.5 mL of 1M TRIS-HCl, pH 8.0
-|50 mM TRIS-HCl
+|25 mM TRIS-HCl
 |-
-|3. Add 30mL of 5M NaCl
+|3. Add 15 mL of 5M NaCl
 |150 mM NaCl
 |-
-|4. Add 2 mL of 0.5M EDTA
+|4. Add 500 μL of 0.5M EDTA
-|1 mM EDTA
+|0.5 mM EDTA
 |-
-|5. Add 100 μL 100% Triton X-100
+|5. Add 5 mL 10% NP-40
-|0.01% Triton X-100
+|0.1% NP-40
 |-
-|5. Bring volume to 1L with millipore H<sub>2</sub>O in a graduated cylinder
+|5. Bring volume to 500L with millipore H<sub>2</sub>O in a graduated cylinder
 |Volume to 1L
 |-
-|6. Immediately prior to use, complete with DTT
+|6. Immediately prior to use, complete with 1μL of 1M DTT per mL TEV CB
 |1 mM DTT
 |}
 [[Category:Yu Solutions]]