How can I do that? I cannot do a single step as there is a too long primer ( with a long hanging tail) and I need to have a blunt end:(
Please any suggestions?
0
3 replies to this topic
#2
scolix
-
- Global Moderators
-
- 310 posts
a student
4
Neutral
Neutral
Posted 28 February 2009 - 08:15 AM
ups, on Feb 27 2009, 12:01 PM, said:
How can I do that? I cannot do a single step as there is a too long primer ( with a long hanging tail) and I need to have a blunt end:(
Please any suggestions?
Please any suggestions?
What is the size of your primer and what is the size of the DNA to be ampified?
#3
ups
-
- Active Members
-
- 12 posts
member
0
Neutral
Neutral
Posted 02 March 2009 - 03:20 AM
The primer is about 70 nucleotides and the fragment that I want to amplify is 3000...
#4
WHR
-
- Active Members
-
- 49 posts
Enthusiast
0
Neutral
Neutral
Posted 02 March 2009 - 07:15 AM
ups, on Mar 2 2009, 03:20 AM, said:
The primer is about 70 nucleotides and the fragment that I want to amplify is 3000...
Hi,
Assume that 20 bases of the 70 is complementary to the template, you may use lower annealing temperature for the first 5 cycles to allow primer binding, and higher temperature for the rest of cycles.
Edited by WHR, 02 March 2009 - 07:15 AM.
