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TruSeq RNA Sample Prep Kit v2 Translated Into Plain English

TruSeq RNA Sample Prep Kit v2 Translated Into Plain English

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2 Comments
  1. Hi Ethan,

    I note in your Plain English protocol that you’ve reduced the bead air-drying time to 2 min. from Illumina’s 15 min. – is this shortened drying time tailored to your particular lab weather conditions, or have you found that 2 min. is long enough, no matter the lab atmosphere, and 15 min. is over the top?

    Aaron.

  2. ethanomics permalink

    Any old molecular biologists will tell you (unless he/she is too old and goes back to the SpeedVac days), drying your nucleic acids makes them harder to get back into solution. I would go as far to say, do not dry your beads at all and especially don’t let them get so dry they crack. The two minutes is more the amount of time it takes to process the samples, so there really is no incubation time. This is confirmed in the Beckman Coulter AMPure XP protocol. I think the molecular biologists at Illumina are a little clueless about some things. Trace amounts of ethanol do not effect any of the downstream reactions. Lab humidity would effect things at all. Just make sure you are careful to remove all the ethanol you can. I go through one time and then go through the samples a second time to get the last microliter of ethanol.

    That and it saves time as well.

    or maybe they are not clueless at Illumina but just assume people won’t aspirate all the ethanol.

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