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!
Photo
- - - - -

difference between cDNA and mRNA


  • Please log in to reply
5 replies to this topic

#1 helpwithdna

helpwithdna

    member

  • Active Members
  • Pip
  • 14 posts
0
Neutral

Posted 22 September 2013 - 11:13 PM

Dear all,

I have to clone a gene which shows "....mRNA, complete cds" on PUBMED.

Now I need to find out the target sequence and design a primer for cloning this gene.

I know I would have to extract the RNA and convert it to cDNA during isolation.

However, can I directly use this "....mRNA, complete cds" to pick primers in blast? Or do I need to pick primers in the mRNA sequence?

Thank you very much for your help.


Edited by helpwithdna, 22 September 2013 - 11:14 PM.


#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,663 posts
394
Excellent

Posted 22 September 2013 - 11:54 PM

Basically the sequence you see on pubmed is the cDNA sequence - RNA would have uracil (U) in the place of thymidine (T), and isn't currently sequenceable in the manner that DNA is. 

 

A forward primer would be the same as the sequence you are seeing on pubmed, and the reverse would be the reverse complement of the sequence you see.



#3 helpwithdna

helpwithdna

    member

  • Active Members
  • Pip
  • 14 posts
0
Neutral

Posted 23 September 2013 - 03:05 AM

Basically the sequence you see on pubmed is the cDNA sequence - RNA would have uracil (U) in the place of thymidine (T), and isn't currently sequenceable in the manner that DNA is. 

 

A forward primer would be the same as the sequence you are seeing on pubmed, and the reverse would be the reverse complement of the sequence you see.

Thanks! I would like to ask, if my forward and reverse primer includes the cds region of the cDNA sequence, is it correct?


Edited by helpwithdna, 23 September 2013 - 03:05 AM.


#4 labbey2.0

labbey2.0

    member

  • Active Members
  • Pip
  • 6 posts
0
Neutral

Posted 30 September 2013 - 08:57 AM

Just remember that cDNA has the intronic sequences removed.



#5 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,663 posts
394
Excellent

Posted 30 September 2013 - 11:42 PM

 

Basically the sequence you see on pubmed is the cDNA sequence - RNA would have uracil (U) in the place of thymidine (T), and isn't currently sequenceable in the manner that DNA is. 

 

A forward primer would be the same as the sequence you are seeing on pubmed, and the reverse would be the reverse complement of the sequence you see.

Thanks! I would like to ask, if my forward and reverse primer includes the cds region of the cDNA sequence, is it correct?

 

Correct for what purpose?  If you want to clone the coding sequence, your primers essentially need to incorporate the start and stop codons, but if you just want to measure presence/absence then any primer pair should work.



#6 labbey2.0

labbey2.0

    member

  • Active Members
  • Pip
  • 6 posts
0
Neutral

Posted 01 October 2013 - 06:55 AM

The only caveat to that is whether your gene is subject to intragenic duplication or full deletion.  But that's usually rare and specific to certain genes.






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

©1999-2013 Protocol Online, All rights reserved.