Yu:TEV Cleavage Buffer

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{| border="0" cellpadding="10" width="100%"
{| border="0" cellpadding="10" width="100%"
|+'''Solution recipe for 1L TEV Cleavage Buffer.'''
|+'''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"|
|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
|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
|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
|1 mM DTT
|}
|}
[[Category:Yu Solutions]]
[[Category:Yu Solutions]]

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
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