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Low 260/230 ratio in my RNA


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#1 Blackeyed

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Posted 11 December 2009 - 03:26 AM

I`ve isolated RNA with the Promega kit. I measured the concentration etc. with Nanodrop and I have low
260/230 ratio`s ( around 1.4-1.8). The troubleshooting section of the kit
said that there is to much guanidine thiocyanate and that I should precipitate my RNA with NaCl and ethanol
at -20 for 30min.
I already have a low concentration of RNA so I`m a bit susceptible to do this because it eventually again
will decrease my RNA concentration.
Will this guanidine thiocyanate comprimise my RT-PCR results? Should I leave my samples
the way they are or should i perform the additional step and probably loose RNA?

#2 gogreen

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Posted 13 December 2009 - 09:44 AM

hey black eyed, a 260/230 of 1.4-1.8 is good enough for the downstream applications...I would go ahead with the next step if I were you.

#3 sanjiun

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Posted 13 December 2009 - 08:20 PM

I guess it should be all right if you constantly use RNA with this range of reading.
I mean it is not like you are using very good quality RNA and quite bad quality RNA sample, and comparing transcription of gene between them. Then you'll have variation problem.




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