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We are extracting RNA from single insects with the later purpose of performing RNA-seq. We are using the RNeasy Plus Miniki from Qiagen, which involves the use of eliminator columns as you can see here page 7.

However, we would love to be able to recover some DNA to perform a PCR which would help us confirm the insect species we are dealing with. Ideally, we would like to do this without losing any RNA-containing material. We were therefore wondering whether it was possible to retrieve some DNA from a column, which would otherwise be discarded ? Does anyone have experience with this ?

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  • $\begingroup$ Have you looked to see if there is DNA in your RNA extraction? $\endgroup$
    – bob1
    Jan 23 at 2:34
  • $\begingroup$ We have run it on a gel yes, there was none after the DNA removal step. $\endgroup$
    – CaroZ
    Jan 23 at 7:42
  • $\begingroup$ Test it with a PCR; gels are about the least sensitive method of looking for DNA. almost every RNA extraction contains some DNA. So long as you didn't add DNase that is. $\endgroup$
    – bob1
    Jan 23 at 20:09
  • $\begingroup$ The idea however is that we would use material which would be otherwise discarded, we are reluctant to take away some of our pool of RNA. $\endgroup$
    – CaroZ
    Jan 25 at 10:09
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    $\begingroup$ Simple - don't use the eliminator columns... Note that most RNAseq preps use in the fmol range of sample loaded and have a PCR amplification step to help you get there. $\endgroup$
    – bob1
    Jan 26 at 20:27

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