In my haste to finish genotyping I added Blue Juice to my PCR product as if I were going to run a gel to check the product... but really I just wanted to digest the product (got stuck on autopilot).
Can I still digest the wells I added blue juice to? Or will I need to re-PCR that DNA?
Thanks much
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3 replies to this topic
#2
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Posted 20 November 2009 - 12:03 PM
Hi alwaysleaning,
BlueJuice, unlike other loading dyes, does not have glycerol (which inhibits restriction enzymes), but instead uses sucrose, which in theory should not inhibit your restriction digest.
My suggestion: if you are in a big rush, try to digest and see what happens (it may work). If not, run the PCRs again.
Good luck
BlueJuice, unlike other loading dyes, does not have glycerol (which inhibits restriction enzymes), but instead uses sucrose, which in theory should not inhibit your restriction digest.
My suggestion: if you are in a big rush, try to digest and see what happens (it may work). If not, run the PCRs again.
Good luck
Ivan
Carlsbad, CA
#3
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Sunflower
Posted 20 November 2009 - 03:21 PM
why re-pcr? just do an EtOH precipitation. clean, fast, easy.
"it is a miracle that curiosity survives formal education" -A.E.
#4
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Enthusiast
Posted 22 November 2009 - 02:35 AM
Another option: Purify the mixture using PCR purification spin-columns.
If you don't know it, then ask it! Better to ask and look foolish to some then not ask and stay stupid.
