Biology Forum › Molecular Biology › Digest
- AuthorPosts
- November 11, 2007 at 8:15 pm #8559
benedictine
Participanthi please may I ask; I did a digest using TaqI; its supposed to have 1,2 or 3 bands according to the polymorphism, but why am I getting sometimes 4 bands and 5. What happened and what should I do to avoid this.
Thank you, from bene - November 15, 2007 at 6:52 am #77788
pluto
Participantdoes it have star activity? use less enzyme next time if so.
- November 15, 2007 at 2:17 pm #77793
benedictine
Participantsorry I dont know what is star activity. What is that?
All I know is it is supposed to cut the genes of alleles of a CYP2 that I’m studying. thanks, bene.also, do you have any idea what to do with acrylamide that has solidified in the beaker (in the refrigerator)? How can we remove it? (I will post this qustion again as a new topic).
thanks again
bene - November 15, 2007 at 3:00 pm #77797
blcr11
ParticipantI usually think of TaqI as a polymerase, so I guess this must be TaqaI? Most likely the extra bands are coming from partial digestion of the restriction sites. TaqaI is sensitive to methylation so if your sites are partially methylated for some reason you may also be getting what looks like partial digestion. I don’t think TaqaI has star activity. Some enzymes, when used at high concentration, can recognize additional sites. This extra activity is called "star" activity and the extra recognition sequence(s) is typically indicated with an astersick, hence the "star" designation.
- November 15, 2007 at 5:23 pm #77799
blcr11
ParticipantThat should’ve been "asterisk," of course.
- November 16, 2007 at 3:53 pm #77853
benedictine
Participantthanks for writing. It says on the bottle TaqI, not Taqal. Thanks a lot. I use at the most 1ul of enzyme per sample, is that a lot? and from where do we check if our sites are partially methylated? is it from sequence?
thanks again, bene - November 16, 2007 at 5:43 pm #77860
canalon
ParticipantI found this enzyme as Taq I with NEB, but it is "Taqa1" on the page:
http://www.neb.ca/detail.lasso?id=R0149&ven=NEB
If you look at the end there is FAQ list, maybe that will be useful, and it says that taq I has some star activity… - November 26, 2007 at 11:01 pm #78526
benedictine
Participantthanks for the link
from bene
- AuthorPosts
You must be logged in to reply to this topic.