CTR:Recipes: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
Sirena Lao (talk | contribs) No edit summary |
Sirena Lao (talk | contribs) No edit summary |
||
| (5 intermediate revisions by the same user not shown) | |||
| Line 1: | Line 1: | ||
Lab recipes for general-use buffers. Also refer to the index card box located near the gel rigs. | 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:Protocols Lab Protocols]</div> | |||
==Queen's Lysis Buffer== | ==Queen's Lysis Buffer== | ||
| Line 7: | Line 11: | ||
*10.0g N-lauroylsarcosine | *10.0g N-lauroylsarcosine | ||
*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> | |||
==TAE Buffer (50x)== | ==TAE Buffer (50x)== | ||
| Line 13: | Line 20: | ||
*57.1mL glacial acetic acid | *57.1mL glacial acetic acid | ||
*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> | |||
==TBE Buffer (5x)== | ==TBE Buffer (5x)== | ||
| Line 19: | Line 29: | ||
*3.72g EDTA (or 20mL 0.5M EDTA pH 8.0) | *3.72g EDTA (or 20mL 0.5M EDTA pH 8.0) | ||
*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> | |||
==TE Buffer== | ==TE Buffer== | ||
*10mM | *10mM TrisHCl of desired pH | ||
*1mM EDTA pH 8.0 | *1mM EDTA pH 8.0 | ||
<div align="right">[http://openwetware.org/wiki/CTR:Recipes Top]</div> | |||
==LowTE Buffer== | ==LowTE Buffer== | ||
| Line 32: | Line 48: | ||
*3μL EDTA 0.5M | *3μL EDTA 0.5M | ||
*14.85mL dH<sub>2</sub>O | *14.85mL dH<sub>2</sub>O | ||
<div align="right">[http://openwetware.org/wiki/CTR:Recipes Top]</div> | |||
==LB Agar Plates== | |||
1. Mix: | |||
*10g Tryptone | |||
*5g Yeast extract | |||
*10g NaCl | |||
*950mL dH<sub>2</sub>O | |||
*15 g/L agar | |||
2. Adjust pH to 7.0 using NaOH and bring volume to 1 L.<br> | |||
3. Autoclave on liquid cycle for 20 minutes at 15 psi.<br> | |||
4. Allow solution to cool to 55°C and add 50 μg/mL of ampicillin or kanamycin.<br> | |||
5. Pour into plates, sterilizing the plate and flask/bottle before each pour.<br> | |||
6. Let plates harden.<br> | |||
7. Store in cold room. | |||
<div align="right">[http://openwetware.org/wiki/CTR:Recipes Top]</div> | |||
Revision as of 15:44, 28 July 2014
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
- 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.
