CTR:Recipes: Difference between revisions
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Lab recipes for general-use buffers. Remember to autoclave! Also refer to the index card box located near the gel rigs. | |||
<div align="right">[http://openwetware.org/wiki/CTR Home]</div> | <div align="right">[http://openwetware.org/wiki/CTR Home]</div> | ||
<div align="right">[http://openwetware.org/wiki/CTR:Protocols Lab Protocols]</div> | <div align="right">[http://openwetware.org/wiki/CTR:Protocols Lab Protocols]</div> | ||
==Queen's Lysis Buffer== | ==Queen's Lysis Buffer== | ||
| Line 8: | Line 9: | ||
*0.58g NaCl | *0.58g NaCl | ||
*3.73g EDTA | *3.73g EDTA | ||
*10.0g N-lauroylsarcosine | *10.0g N-lauroylsarcosine (a more potent version of the cell lysing agent (SDS) recommended by the USDA - 10%) | ||
*dH<sub>2</sub>O to 1L total volume | *dH<sub>2</sub>O to 1L total volume | ||
<div align="right">[http://openwetware.org/wiki/CTR:Recipes Top]</div> | <div align="right">[http://openwetware.org/wiki/CTR:Recipes Top]</div> | ||
==TAE Buffer (50x)== | ==TAE Buffer (50x)== | ||
Latest revision as of 15:53, 27 June 2016
Lab recipes for general-use buffers. Remember to autoclave! Also refer to the index card box located near the gel rigs.
Queen's Lysis Buffer
- 1.21g Tris Base
- 0.58g NaCl
- 3.73g EDTA
- 10.0g N-lauroylsarcosine (a more potent version of the cell lysing agent (SDS) recommended by the USDA - 10%)
- dH2O to 1L total volume
TAE Buffer (50x)
- 242g Tris Base
- 23.8g EDTA (or 100mL 0.5M EDTA pH 8.0)
- 57.1mL glacial acetic acid
- dH2O to 1L total volume
TBE Buffer (5x)
- 54g Tris Base
- 27.5g Boric acid
- 3.72g EDTA (or 20mL 0.5M EDTA pH 8.0)
- dH2O to 1L total volume
TE Buffer
- 10mM TrisHCl of desired pH
- 1mM EDTA pH 8.0
LowTE Buffer
- 10mM TrisHCl
- 0.1mM EDTA pH 8.0
example to make 15mL:
- 150μL TrisHCl 1M
- 3μL EDTA 0.5M
- 14.85mL dH2O
LB Agar Plates
1. Mix:
- 10g Tryptone
- 5g Yeast extract
- 10g NaCl
- 950mL dH2O
- 15 g/L agar
2. Adjust pH to 7.0 using NaOH and bring volume to 1 L.
3. Autoclave on liquid cycle for 20 minutes at 15 psi.
4. Allow solution to cool to 55°C and add 50 μg/mL of ampicillin or kanamycin.
5. Pour into plates, sterilizing the plate and flask/bottle before each pour.
6. Let plates harden.
7. Store in cold room.
