ARE THERE ANY GENETICS STUDENTS HERE? RNA IS KILLING ME

N

help! RNA extraction is killing me.

S

what protocol are you using? what are u extracting from?

N

I'm using Ribopure kit(see link below). I'm extracting RNA for microarrays in yeast, and I'm really annoyed, because I finaly got 40micrograms, but even though the A(260/280) is 2.1, I keep getting A(260/230) around 0.22! I've got a huge peak on the 230, and I cant work out wat it is. I avoided the phenol, and I have a pretty clean space, I'm starting to think the last DNAse treatment step is ruining it for me!:-s
how on earth do i get rid of that peak. an ex lab member used to get clean and I cannot get a hold of them(PI is busy of course) and I have noone else to ask. thanks for your reply!

http://www.ambion.com/techlib/prot/fm_1926.pdf

N

Quote From satchi:

what protocol are you using? what are u extracting from?

S

do u think maybe the DNAse didnt cut properly; also could it be that there is a contamination problem somewhere.
Are there any lab members who can help you out? like you could go to the next lab and ask.

also there is an RNA forum here you can ask.
http://www.molecularstation.com/forum/

N

Quote From satchi:

do u think maybe the DNAse didnt cut properly; also could it be that there is a contamination problem somewhere.
Are there any lab members who can help you out? like you could go to the next lab and ask.

also there is an RNA forum here you can ask.
http://www.molecularstation.com/forum/



thank you so much for your help, turns out, since my yield was good, the 260/280 was ethanol for the column. in the trouble shooting section it emphasises to CAREFULLY remove the column from the collection tube.



best wishes!

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