Oneill Lab:Enzymes: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
No edit summary |
No edit summary |
||
Line 26: | Line 26: | ||
Aliquot and store at -20°C | Aliquot and store at -20°C | ||
===Nuclease Solution=== | |||
<small>Used in lambda extractions prior to capsid degradation.</small> | |||
*DNaseI 10mg | |||
*RNaseA 10mg | |||
*1M Tris 10μl | |||
*5M NaCl 30μl | |||
*1M MgCl<sub>2</sub> 1μl | |||
*Water 950μl | |||
Use at concentration of 1mg/ml | |||
==Links and Contacts== | ==Links and Contacts== |
Latest revision as of 21:21, 7 May 2007
Michael J. O'Neill Lab University of Connecticut Department of Molecular and Cell Biology
Lab Stock Enzymes
Preparation of lab stock enzymes and buffers
10x EcoRI Buffer
- Combine
- 8mL 2.5M Tris
- 2mL 5M NaCl
- 1mL 1M MgCl2
- 50μL TritonX-100
- Adjust pH to 7.5 with 12M HCl
- Add water to 20mL
Proteinase K
Makes 0.5mL at 20mg/ml
- 10μL 1M CaCl2
- 5μL 1M Tris pH 7.5
- 250μL Glycerol
- Pipette with cut tip
- Use under flame
- 10mg ProK
- Stored in 4°C dessicator
- 235μL H2O
Aliquot and store at -20°C
Nuclease Solution
Used in lambda extractions prior to capsid degradation.
- DNaseI 10mg
- RNaseA 10mg
- 1M Tris 10μl
- 5M NaCl 30μl
- 1M MgCl2 1μl
- Water 950μl
Use at concentration of 1mg/ml