A couple of things
1) Nested enzymes
There is no such thing…but there are such things as nested restriction site recognition sites and molecular biologists who make up confusing expressions.
The idea is that,sometimes, when you cut a vector with enzyme I and the DNA to be inserted with enzyme II, and you join them together, you may re-generate the recognition site for enzyme I or enzyme II (depending on the situation) or generate a site for another enzyme altogether (enzyme III). In some cases, the outcome of the rejoining procedure depends on what nucleotide is present in the insert just adjacent to the restriciton site for enzyme II.
What may be confusing is that you may be cutting the vector with enzyme I, cutting the genomic DNA with enzyme II, and after rejoining you obtain 3 clones of interest (containing different genomic fragments), and clone I may have regenerated one site for enzyme I at one junction, while clone II may not have regenerated any site, and clone 3 may have regenerated the site for enzyme I at both junctions.
Just treat each clone as a separate entity!
2) Drugs and cell signalling : here’s a fun site if you need to take a short break from studying: go to
http://learn.genetics.utah.edu/units/addiction/drugs/mouse.cfm
3) Please remember : you won’t need to memorize any gene names or functions. Just keep in mond some examples of mechanisms, e.g. gradients of proteins/RNAs, cell-to-cell signalling, transcriptional regulation, post-transcriptional regulation (e.g. inhibition of translation), etc.
4) tomorrow is review day. Try to come in with a list of specific questions that you have....
There is no such thing…but there are such things as nested restriction site recognition sites and molecular biologists who make up confusing expressions.
The idea is that,sometimes, when you cut a vector with enzyme I and the DNA to be inserted with enzyme II, and you join them together, you may re-generate the recognition site for enzyme I or enzyme II (depending on the situation) or generate a site for another enzyme altogether (enzyme III). In some cases, the outcome of the rejoining procedure depends on what nucleotide is present in the insert just adjacent to the restriciton site for enzyme II.
What may be confusing is that you may be cutting the vector with enzyme I, cutting the genomic DNA with enzyme II, and after rejoining you obtain 3 clones of interest (containing different genomic fragments), and clone I may have regenerated one site for enzyme I at one junction, while clone II may not have regenerated any site, and clone 3 may have regenerated the site for enzyme I at both junctions.
Just treat each clone as a separate entity!
2) Drugs and cell signalling : here’s a fun site if you need to take a short break from studying: go to
http://learn.genetics.utah.edu/units/addiction/drugs/mouse.cfm
3) Please remember : you won’t need to memorize any gene names or functions. Just keep in mond some examples of mechanisms, e.g. gradients of proteins/RNAs, cell-to-cell signalling, transcriptional regulation, post-transcriptional regulation (e.g. inhibition of translation), etc.
4) tomorrow is review day. Try to come in with a list of specific questions that you have....
posted by pam39 at 4:57 PM
6 Comments:
PAm, can we go over some of the developmetal type questions we may see on the final tomorrow.
Also, do you have more of those RFLP and VNTR q's that we could work on and possibly some more of the transposable elements. Thanks again and I really hope I haven't overwhelmed you with all the question requests.
There are some good practice problems on the main 335 web page!
I'll try to make one up with VNTRs
Cheers
Pam
i like that animation~
Pam, you know the developmental q's on the 335 website, are we supposed to be able to do those? I have no idea how to go about them!!!!
with VNTR we have one allele and one VNTR locus, with RFLP we have one allele, but multiple loci?
I don't know if any of you are going to check this blog..but what do you guys think about the final exam?
I thought it was hard... very hard (except for Q2)
Q4,7 were really hard. I didin't really know how to do Q7...
I just hope that I passed the exam.
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