QUESTIONS THAT CAME UP!
Thank you very much for taking the time to ask all these questions in an organized way. I tried to do my best to clarify...let me know if it helps (or if it doesn't!)
Cheer,
Pam
posted by pam39 at 8:58 PM
Tuesday, July 03, 2007
Thank you very much for taking the time to ask all these questions in an organized way. I tried to do my best to clarify...let me know if it helps (or if it doesn't!)
Cheer,
Pam
BIOL335 links
- Your textbook!
- UBC BIOL335 website(updated)
- More than you'll need to know on P elements
- Link to a site on Ac/Ds elements
- Landmark discoveries in molecular biology
- Link to mol.gen. of prokaryotes course at univ. of Arizona
- Lac operon stuff
- Link to a clatech problem set
- Link to another caltech problem set
- Link to Dr. Zimmerman's molbiol-check out their links!
- BIOL334 website
Genetics links
- Your textbook!
- Traditional genetics terms
- Human genetics problem sets (University of Arizona)
- Mixed problem set
- Another problem set (vcu)
- Another problem set (vcu)
- Pam's loppins questions
- Triploid bananas: how does it work?
- BIOL334 website
Previous Posts
- TECHNIQUUES
- ANNOUNCEMENT
- SOME QUESTIONS THAT CAME UP
- WELCOME TO BIOL335!
- Review and quant genetics
- A few practice questions-up to chapter 15
- ON 'GENE INTERACTION'
- BACTERIA STUFF
- ABOUT TETRADS
- Pre-MT review
10 Comments:
Wow, Pam! You're so fast!!! Thanks!
It helped!
Hi Pam I was just wondering if when you are making a genomic library you have to use the same restriction enzyme? We only use genomic libraries to determine the entire gene sequence and we use cDNA libraries to find the potential sequence since it is smaller and easier to use?
Sorry...you have to give me a few more details. For example "the same RE" ...same as what?
We use genomic libraries for a whole bunch of things. To find/clone genes, to find/clone regulatory sequences, to map things relative to each other, to characterize the structure of a gene, etc.
cDNA libraries are used for various applications, too. Some examples that came up: to clone a gene based on its expression pattern/profile, to find a gene of interest and put it directly into an expression vector, to derive cDNA arrays, etc.
I am not sure what you mean by "potential sequence"...
Cheers,
Pam
By the way, on the post below you should click on "here" to see the techniques page.
I can't seem to download Omid's mock midterm exam. Is anyone having same problem??
I was trying to download it as well and i cannot get access to it either
so does anyone think we will be getting a cloning "project" type of question? Craig said that there will be no ORF stuff, so we're left with recombinant maps and Lac stuff. Can someone just go over the basics of cloning since I am having a horrible time with it
Pam , can we go over some of the practice questions that are on the 335 website, especially #1 from the 2006 midterm.
Thanks so much
Ok. This is what I think what's going to be on the midterm.
1. Prokaryotic gene regulation
e.g. Is there going to be growth if you put X-gal/IPTG/antibiotic, etc.? Is it going to be white/blue/no colony at all??
2. Restriction Mapping
-Hopefully we won't have "nested enzyme" ^0^
3. Cloning Project
-Remember the "Midterm Stress gene" stuff we went over during lecture???
Can someone tell me if this is right please.
Cloning:
1) do a partial digest on DNA using restriction enzyme of choice
2) use the same restriction enzyme as above to do a complete digest on a vector of choice (one with an antibiotic resistance or Lac Z+)
3) next add vector plasmid and donor DNA with ligase to attach them together
4) transform into bacteria that is sensitive to the antibiotic or Lac Z- to the plasmid we chose above
5) plate on a media with the antibiotic, those that recieved the plasid are the ones that will grow on the media, this is now our genomic library (?)
6) colony lift hybridization is used to determine which one of the clones has our gene of interest by running it on a agrose southern blot
7) to determine which one of the clones has the gene we use a probe for the gene along with the genomic library
8) we can then use ethidium bromide or autorad to see the bands that are made by those locations that are similar or identical to the probe thus we have found the gene we were looking for
9)does PCR come next in order to amplify the gene?
I am not sure if these are the steps, it will be great if someone can tell me if I am on going in the right direction or if I have missed the point :)
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