RNA Electrophoresis Hints - (Dec/03/2008 )
Hi again, I previously asked about QC in RNA purification due to poor purity index A260/A230. I got over this trouble and I'm currently extracting RNA in goog amount and quality.
The next step is to check RNA integrity by running it in denaturant gel electrophoresis... and I have a question: (I use a formamide/denaturing agarose gel electrophoresis protocol)
¿Should I perform it under RNAase-free conditions? I mean, should all components (buffers, agarose, formamide, ...) be RNAase free? RNAases can digest my sample mixtures (RNA + Formamide + Ethiduim bromide + dye/glicerol) inside the gel???
How do you perform?
Finaly, there were three questions. Thanks in advance!!!
-Luis SC-
QUOTE (Luis SC @ Dec 3 2008, 07:03 PM)
Hi again, I previously asked about QC in RNA purification due to poor purity index A260/A230. I got over this trouble and I'm currently extracting RNA in goog amount and quality.
The next step is to check RNA integrity by running it in denaturant gel electrophoresis... and I have a question: (I use a formamide/denaturing agarose gel electrophoresis protocol)
¿Should I perform it under RNAase-free conditions? I mean, should all components (buffers, agarose, formamide, ...) be RNAase free? RNAases can digest my sample mixtures (RNA + Formamide + Ethiduim bromide + dye/glicerol) inside the gel???
How do you perform?
Finaly, there were three questions. Thanks in advance!!!
The next step is to check RNA integrity by running it in denaturant gel electrophoresis... and I have a question: (I use a formamide/denaturing agarose gel electrophoresis protocol)
¿Should I perform it under RNAase-free conditions? I mean, should all components (buffers, agarose, formamide, ...) be RNAase free? RNAases can digest my sample mixtures (RNA + Formamide + Ethiduim bromide + dye/glicerol) inside the gel???
How do you perform?
Finaly, there were three questions. Thanks in advance!!!
Yes all water must be DEPC-treated. but RNases generally don't work that fast as to digest all your RNA until nothing can be seen after 45 min of gel run.
-chrisbelle-
QUOTE (Luis SC @ Dec 3 2008, 07:03 PM)
Hi again, I previously asked about QC in RNA purification due to poor purity index A260/A230. I got over this trouble and I'm currently extracting RNA in goog amount and quality.
The next step is to check RNA integrity by running it in denaturant gel electrophoresis... and I have a question: (I use a formamide/denaturing agarose gel electrophoresis protocol)
¿Should I perform it under RNAase-free conditions? I mean, should all components (buffers, agarose, formamide, ...) be RNAase free? RNAases can digest my sample mixtures (RNA + Formamide + Ethiduim bromide + dye/glicerol) inside the gel???
How do you perform?
Finaly, there were three questions. Thanks in advance!!!
The next step is to check RNA integrity by running it in denaturant gel electrophoresis... and I have a question: (I use a formamide/denaturing agarose gel electrophoresis protocol)
¿Should I perform it under RNAase-free conditions? I mean, should all components (buffers, agarose, formamide, ...) be RNAase free? RNAases can digest my sample mixtures (RNA + Formamide + Ethiduim bromide + dye/glicerol) inside the gel???
How do you perform?
Finaly, there were three questions. Thanks in advance!!!
Yes all water must be DEPC-treated. but RNases generally don't work that fast as to digest all your RNA until nothing can be seen after 45 min of gel run. but yes try not to mix DNA and RNA gel tanks, moulds and loading area
-chrisbelle-