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Team:The University of Alberta/17 June 2008
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==Troubleshooting Results== | ==Troubleshooting Results== | ||
'''Saima'''<br> | '''Saima'''<br> | ||
| - | Tried doing digests from Jason's digests of the genes in pUC57 (see below) using columns from three different kits. The results were all similar - very low concentrations for each. Next we are going to try running already purified DNA of a known, high concentration through the columns and measure the concentration that is eluted to determine how much, if any, is sticking to the column | + | Tried doing digests from Jason's digests of the genes in pUC57 (see below) using columns from three different kits. The results were all similar - very low concentrations for each. Next we are going to try running already purified DNA of a known, high concentration through the columns and measure the concentration that is eluted to determine how much, if any, is sticking to the column<br> |
'''Jason'''<br> | '''Jason'''<br> | ||
| - | + | [[Image:june17_puc57_genes.jpg|thumb]] | |
| - | + | *The digests of the reporter genes in pUC57 were run on a 1% agarose gel. The bands were the correct size; see the image to the right. | |
| + | *Making a single cut with Xba in the J61003 vector to attempt to ligate it back together and thereby testing our ligation buffer and enzyme. This will let us know if the reason our transformations havent been working is due to bad ligase buffer/enzyme (eg, maybe the ATP in the ligase buffer is no good anymore). | ||
| + | *also redoing the Purple Russian J61003 ligation using new ligation buffer | ||
| + | '''Chris'''<br> | ||
| + | *Continuing with the westerns. Made some new destain solution which should last for a while. | ||
| + | '''Tom''' | ||
| + | *Made stocks of TetR and I0500 from the iGEM Parts Binder | ||
| + | |||
| + | ==Plant Project== | ||
| + | Our first batch of plants are almost flowering on schedual with the Six week schedual we set for them | ||
| + | A new tray was planted to day and put into the growth chamber | ||
| + | |||
| + | ==David's Crusade== | ||
| + | Ok, so things are not working anywhere near as well as they ought to be working so David is going to take a run at making the Purple Russian, Blue Ox and Tryp biobricks. | ||
| + | |||
| + | The quest begins with the plasmids given to us by Mike's lab and forward and reverse primers for each that include the correct biobrick prefixes and suffixes. | ||
| + | |||
| + | I have amplified each gene using its primers, ''pfu'' proof-reading polymerase and the following pcr program: | ||
| + | |||
| + | 5 min 95C | ||
| + | (30s 95C, 30s 56C, 2 min 72C) x 30 | ||
| + | 5 min 72C | ||
| + | |||
| + | We'll see the results and move forward on June 19th, 2008 | ||
Latest revision as of 23:59, 17 June 2008
Contents |
Today
Today we are continuing the troubleshooting from yesterday.
Troubleshooting Results
Saima
Tried doing digests from Jason's digests of the genes in pUC57 (see below) using columns from three different kits. The results were all similar - very low concentrations for each. Next we are going to try running already purified DNA of a known, high concentration through the columns and measure the concentration that is eluted to determine how much, if any, is sticking to the column
Jason
- The digests of the reporter genes in pUC57 were run on a 1% agarose gel. The bands were the correct size; see the image to the right.
- Making a single cut with Xba in the J61003 vector to attempt to ligate it back together and thereby testing our ligation buffer and enzyme. This will let us know if the reason our transformations havent been working is due to bad ligase buffer/enzyme (eg, maybe the ATP in the ligase buffer is no good anymore).
- also redoing the Purple Russian J61003 ligation using new ligation buffer
Chris
- Continuing with the westerns. Made some new destain solution which should last for a while.
Tom
- Made stocks of TetR and I0500 from the iGEM Parts Binder
Plant Project
Our first batch of plants are almost flowering on schedual with the Six week schedual we set for them A new tray was planted to day and put into the growth chamber
David's Crusade
Ok, so things are not working anywhere near as well as they ought to be working so David is going to take a run at making the Purple Russian, Blue Ox and Tryp biobricks.
The quest begins with the plasmids given to us by Mike's lab and forward and reverse primers for each that include the correct biobrick prefixes and suffixes.
I have amplified each gene using its primers, pfu proof-reading polymerase and the following pcr program:
5 min 95C (30s 95C, 30s 56C, 2 min 72C) x 30 5 min 72C
We'll see the results and move forward on June 19th, 2008
