1,262
edits
In vitro transcription with T7 RNA polymerase: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
In vitro transcription with T7 RNA polymerase (view source)
Revision as of 19:08, 19 June 2005
, 19 June 2005no edit summary
No edit summary |
No edit summary |
||
| Line 66: | Line 66: | ||
Add 5 µL of 500 mM EDTA to stop the reaction. | Add 5 µL of 500 mM EDTA to stop the reaction. | ||
Clean-up/process the RNA | ===Clean-up/process the RNA=== | ||
*[[phenol/chloroform extraction]] followed by [[nucleic acid precipitation]] | *[[phenol/chloroform extraction]] followed by [[nucleic acid precipitation]] | ||
| Line 74: | Line 74: | ||
*Qiagen RNeasy mini kit | *Qiagen RNeasy mini kit | ||
Determine the concentration of your RNA | ===Determine the concentration of your RNA=== | ||
*Always store RNA at a neutral pH with some amount of EDTA. I recommend TE buffer (10 mM Tris-Cl, pH 7.5, 1 mM EDTA). Thinking | * [[Quantification of nucleic acids]] | ||
===Storing RNA=== | |||
*Always store RNA at a neutral pH with some amount of EDTA. I recommend TE buffer (10 mM Tris-Cl, pH 7.5, 1 mM EDTA). Thinking "I'll just put it in water" is a bad idea for RNA (and DNA and proteins and...). Do you really know what's in that water? | |||
Store RNA at -20 ˚C. | Store RNA at -20 ˚C. | ||
