Difference between revisions of "WEEK FOUR (February 6 - 10)"
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| − | We have been given the DNA encoding all | + | We have been given the DNA encoding all 18 genes in 10 different clones and we need to get the DNA into cells, verify the inserts, freeze down glycerol stocks, enter them into [http://gcat.davidson.edu/GCATalog/ GCAT-alog], and send some cells to our colleagues at MWSU. |
[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]. | + | * '''Everyone should register''' for a [http://gcat.davidson.edu/GCATalog/ GCAT-alog account]. |
| + | * The plan this week is to do all wet lab stuff. Dr. Heyer will be out of town and we need to get the magnetasome DNA into ''E. coli'' cells (strain JM109). I know none of you can come to all these meetings, but we will be doing each step as outlined over the week. Come to the ones you can attend. Each person should be able to do at least 3 of these. You can choose which three. | ||
| − | Friday February 3<br> | + | '''Friday February 3 after lab meeting'''<br> |
transform JM109 plate on appropriate media. (Confirm with UW team)<br> | transform JM109 plate on appropriate media. (Confirm with UW team)<br> | ||
take about 30 minutes<br> | take about 30 minutes<br> | ||
| − | Saturday February 4 <br> | + | '''Saturday February 4''' I ''need one volunteer''<br> |
take out of incubator put in fridge<br> | take out of incubator put in fridge<br> | ||
take about 15 minutes and must be done before 9 am<br> | take about 15 minutes and must be done before 9 am<br> | ||
| − | Sunday February 5<br> | + | '''Sunday February 5 late afternoon about 5 pm'''<br> |
pick colonies from each plate and put into 2 mL appropriate media<br> | pick colonies from each plate and put into 2 mL appropriate media<br> | ||
take about 30 minutes<br> | take about 30 minutes<br> | ||
| − | Monday<br> | + | '''Monday February 6 4:30 pm'''<br> |
| − | Do [http://www.bio.davidson.edu/courses/Molbio/Protocols/Zippy_MiniPrep.html minipreps] but SAVE THE CELLS on plates | + | Do [http://www.bio.davidson.edu/courses/Molbio/Protocols/Zippy_MiniPrep.html minipreps] but SAVE THE CELLS on plates<br> |
| − | takes about 1 hour | + | takes about 1 hour<br> |
| − | Tuesday<br> | + | '''Tuesday February 7 4:30 pm'''<br> |
| − | set up [http://www.bio.davidson.edu/courses/Molbio/Protocols/digestion.html digestions], run 30 min to 1 hr and put in fridge | + | set up [http://www.bio.davidson.edu/courses/Molbio/Protocols/digestion.html digestions], run 30 min to 1 hr and put in fridge<br> |
| − | Determine insert sizes from [http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2011&group=Washington Registry] | + | Determine insert sizes from [http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2011&group=Washington Registry]<br> |
| − | Wednesday <br> | + | '''Wednesday February 8 4:30 pm'''<br> |
| − | [http://gcat.davidson.edu/iGEM08/gelwebsite/gelwebsite.html determine the percent agarose] needed for the inserts | + | [http://gcat.davidson.edu/iGEM08/gelwebsite/gelwebsite.html determine the percent agarose] needed for the inserts<br> |
| − | [http://www.bio.davidson.edu/courses/Molbio/Protocols/pourgel.html pour and run gel(s)] then photograph gel(s) | + | [http://www.bio.davidson.edu/courses/Molbio/Protocols/pourgel.html pour and run gel(s)] then photograph gel(s)<br> |
| − | verify insert sizes (get this from [http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2011&group=Washington Registry]) | + | verify insert sizes (get this from [http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2011&group=Washington Registry])<br> |
| − | Thursday<br> | + | '''Thursday February 9 11 am'''<br> |
| − | grow overnight cultures of clones with correct inserts sizes | + | grow overnight cultures of clones with correct inserts sizes<br> |
| − | pick from plate and put into 2 mL of appropriate media | + | pick from plate and put into 2 mL of appropriate media<br> |
| − | Friday <br> | + | '''Friday February 10 4 pm'''<br> |
| − | take cells out of incubator | + | take cells out of incubator<br> |
| − | freeze down cells and enter into [http://gcat.davidson.edu/GCATalog/ GCAT-alog]. | + | freeze down cells and enter into [http://gcat.davidson.edu/GCATalog/ GCAT-alog].<br> |
Revision as of 02:46, 26 January 2012
We have been given the DNA encoding all 18 genes in 10 different clones 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.
- The plan this week is to do all wet lab stuff. Dr. Heyer will be out of town and we need to get the magnetasome DNA into E. coli cells (strain JM109). I know none of you can come to all these meetings, but we will be doing each step as outlined over the week. Come to the ones you can attend. Each person should be able to do at least 3 of these. You can choose which three.
Friday February 3 after lab meeting
transform JM109 plate on appropriate media. (Confirm with UW team)
take about 30 minutes
Saturday February 4 I need one volunteer
take out of incubator put in fridge
take about 15 minutes and must be done before 9 am
Sunday February 5 late afternoon about 5 pm
pick colonies from each plate and put into 2 mL appropriate media
take about 30 minutes
Monday February 6 4:30 pm
Do minipreps but SAVE THE CELLS on plates
takes about 1 hour
Tuesday February 7 4:30 pm
set up digestions, run 30 min to 1 hr and put in fridge
Determine insert sizes from Registry
Wednesday February 8 4:30 pm
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 February 9 11 am
grow overnight cultures of clones with correct inserts sizes
pick from plate and put into 2 mL of appropriate media
Friday February 10 4 pm
take cells out of incubator
freeze down cells and enter into GCAT-alog.
