Difference between revisions of "WEEK FOUR (February 6 - 10)"
Macampbell (talk | contribs) (Created page with 'We have been given the DNA encoding all 11 genes and we need to get the DNA into cells, verify the inserts, freeze down glycerol stocks, enter them into [http://gcat.davidson.edu…') |
Macampbell (talk | contribs) |
||
| Line 2: | Line 2: | ||
[http://gcat.davidson.edu/mediawiki-1.15.0/index.php/Davidson_Protocols Davidson Protocols] | [http://gcat.davidson.edu/mediawiki-1.15.0/index.php/Davidson_Protocols Davidson Protocols] | ||
| + | |||
| + | |||
| + | Everyone should register for a [http://gcat.davidson.edu/GCATalog/ GCAT-alog account]. | ||
| + | |||
| + | Friday February 3<br> | ||
| + | transform JM109 plate on appropriate media. (Confirm with UW team)<br> | ||
| + | take about 30 minutes<br> | ||
| + | |||
| + | |||
| + | Saturday February 4 <br> | ||
| + | take out of incubator put in fridge<br> | ||
| + | take about 15 minutes and must be done before 9 am<br> | ||
| + | |||
| + | |||
| + | Sunday February 5<br> | ||
| + | pick colonies from each plate and put into 2 mL appropriate media<br> | ||
| + | take about 30 minutes<br> | ||
| + | |||
| + | |||
| + | Monday<br> | ||
| + | Do [http://www.bio.davidson.edu/courses/Molbio/Protocols/Zippy_MiniPrep.html minipreps] but SAVE THE CELLS on plates | ||
| + | takes about 1 hour | ||
| + | |||
| + | |||
| + | Tuesday<br> | ||
| + | set up [http://www.bio.davidson.edu/courses/Molbio/Protocols/digestion.html digestions], run 30 min to 1 hr and put in fridge | ||
| + | Determine insert sizes from [http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2011&group=Washington Registry] | ||
| + | |||
| + | |||
| + | Wednesday <br> | ||
| + | [http://gcat.davidson.edu/iGEM08/gelwebsite/gelwebsite.html determine the percent agarose] needed for the inserts | ||
| + | [http://www.bio.davidson.edu/courses/Molbio/Protocols/pourgel.html pour and run gel(s)] then photograph gel(s) | ||
| + | verify insert sizes (get this from [http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2011&group=Washington Registry]) | ||
| + | |||
| + | |||
| + | Thursday<br> | ||
| + | grow overnight cultures of clones with correct inserts sizes | ||
| + | pick from plate and put into 2 mL of appropriate media | ||
| + | |||
| + | |||
| + | Friday <br> | ||
| + | take cells out of incubator | ||
| + | freeze down cells and enter into [http://gcat.davidson.edu/GCATalog/ GCAT-alog]. | ||
Revision as of 02:35, 26 January 2012
We have been given the DNA encoding all 11 genes and we need to get the DNA into cells, verify the inserts, freeze down glycerol stocks, enter them into GCAT-alog, and send some cells to our colleagues at MWSU.
Everyone should register for a GCAT-alog account.
Friday February 3
transform JM109 plate on appropriate media. (Confirm with UW team)
take about 30 minutes
Saturday February 4
take out of incubator put in fridge
take about 15 minutes and must be done before 9 am
Sunday February 5
pick colonies from each plate and put into 2 mL appropriate media
take about 30 minutes
Monday
Do minipreps but SAVE THE CELLS on plates
takes about 1 hour
Tuesday
set up digestions, run 30 min to 1 hr and put in fridge
Determine insert sizes from Registry
Wednesday
determine the percent agarose needed for the inserts
pour and run gel(s) then photograph gel(s)
verify insert sizes (get this from Registry)
Thursday
grow overnight cultures of clones with correct inserts sizes
pick from plate and put into 2 mL of appropriate media
Friday
take cells out of incubator
freeze down cells and enter into GCAT-alog.
