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RNA extraction: low RCF by mistake


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#1 SF_HK

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Posted 03 January 2013 - 01:58 AM

Hi,

I hope someone's made a similar mistake as me and can guide/assure me.

I had mistakenly done RNA extraction using just 1100 RCF instead of 11,000 RCF. I realised by mistake after treating with isopropanol and centrifugation at 1100 RCF. The last step, which requires washing with ethanol was conducted at the right RCF, i.e. 7500 RCF. I did see a RNA pellet and my RNA purity: 260/280: 1.96, 260/263: 1.68



Thanks

#2 David C H

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Posted 03 January 2013 - 08:07 AM

If you have enough RNA and it looks clean, proceed. The only problem that may result from a short, low-speed spin is that your yield may be low. The spin doesn't do anything special, just forces your RNA to pellet at the bottom of the tube. You may not have spun the sample long enough or fast enough to pellet all of the RNA.




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