Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
- - - - -

simple pcr questions

  • Please log in to reply
1 reply to this topic

#1 vwracer06



  • Active Members
  • Pip
  • 14 posts

Posted 02 October 2002 - 11:08 PM

Hi. Need help on some really basic pcr steps.

Once I have my 50ul reaction micture, how much of that do i put in my actual pcr reaction tubes (the ones that go in the cycler)?

Also what is difference between DEPC and deionized water?

Also, once i have my pcr cycler set up and running, when do I start making the agarose gel? Can i start right away and let it solidify or is it not good to make it too early and let it sit?

Finally, what is a general note or rule for working with chemicals, should i always try to keep everything in the ice bucket?


#2 peterj



  • Members
  • Pip
  • 2 posts

Posted 15 October 2002 - 11:22 PM


You can put all of it in a reaction tube (some use 25ul, other 100ul). I always use 50ul.

DEPC=di-ethylpyrocarbonate (destroys RNases) --> used when working with RNA to prevent degradation

You even can make the gel the day before as long as you put it in the running buffer so that it doesn't dry out

Yes, especially the polymerase. You best divide the NTPs in different aliquots so that you don't have to defreeze and freeze the hole amount each time


Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.