Hi guys,
this is my first thread here, and really appreciate any input from you. Sorry for posting this again here. It seems to fit here better than in sequencing techniques part.
1)
I wonder how you guys titrate adapter to match DNA material amount before ligation? (SE library construction of illumina)
Coz the volume at this stage is only 10ul and the total amount of DNA after many steps of purification is probably only a couple ng. At this low concentration, how do you guys manage to titrate?
2)
We noticed varying amount of fragments at size around 58bp and 82bp in our illumina library. What do you guys think these are? People have been saying that adapter dimer is around 120bp and primer dimer 80-85bp. But how come adapter dimer being 120bp, with each adapter being only 33-34bp? And for primer dimer, 58bp(primer 1.1)+34bp(primer 2.1) is not 80-85bp either.
Thanks for your help!
-xzhang
this is my first thread here, and really appreciate any input from you. Sorry for posting this again here. It seems to fit here better than in sequencing techniques part.
1)
I wonder how you guys titrate adapter to match DNA material amount before ligation? (SE library construction of illumina)
Coz the volume at this stage is only 10ul and the total amount of DNA after many steps of purification is probably only a couple ng. At this low concentration, how do you guys manage to titrate?
2)
We noticed varying amount of fragments at size around 58bp and 82bp in our illumina library. What do you guys think these are? People have been saying that adapter dimer is around 120bp and primer dimer 80-85bp. But how come adapter dimer being 120bp, with each adapter being only 33-34bp? And for primer dimer, 58bp(primer 1.1)+34bp(primer 2.1) is not 80-85bp either.
Thanks for your help!
-xzhang