Team:Technion/21 September 2012
From 2012.igem.org
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-I digested In1 with PstI and XmaI to verify if the clone is OK. After running on gel, I found out that the polymerase is missing in the part- Now I have to begin the cloning all over again! <br> | -I digested In1 with PstI and XmaI to verify if the clone is OK. After running on gel, I found out that the polymerase is missing in the part- Now I have to begin the cloning all over again! <br> | ||
- Restriction of pSB1C3 with XbaI- for my next steps of cloning the parts to the shipping plasmid.<br> | - Restriction of pSB1C3 with XbaI- for my next steps of cloning the parts to the shipping plasmid.<br> | ||
- | -I made 10 starters for In1 from the transformation plates- I picked them randomly.. 1 starter for T7*RNAP+pSB1AK3 (that I know it works) and 4 starters of native T3 RNAP+pSB1AK3 from the transformation plates (picked by random). | + | -I made 10 starters for In1 from the transformation plates- I picked them randomly.. 1 starter for T7*RNAP+pSB1AK3 (that I know it works) and 4 starters of native T3 RNAP+pSB1AK3 from the transformation plates (picked by random).<br> |
+ | -Colony PCR for 2 colonies of In1 part, I stored them at 16C at the end of the PCR. | ||
==Asaf== | ==Asaf== | ||
I got clones from all the transformations including my negative control. I descided due to lack of time<br> | I got clones from all the transformations including my negative control. I descided due to lack of time<br> |
Revision as of 16:08, 22 September 2012
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Ilya
- Wiki, wiki, wiki....
Inbal
-Glycerol stocks for In1+ALP and In1+xyIE.
-Also, we have done the assay for the In1+ALP and In1+xyIE, but the results were the same as the control or much lesser than the control..... :/ at my opinion- I missed the eT7 RNAP somewhere in the cloning... Also, we checked the mCherry fluorescence, which is very high compare to the control- so I'm sure that the promoter with the mCherry is within the plasmid (pSB1AK3).
-I digested In1 with PstI and XmaI to verify if the clone is OK. After running on gel, I found out that the polymerase is missing in the part- Now I have to begin the cloning all over again!
- Restriction of pSB1C3 with XbaI- for my next steps of cloning the parts to the shipping plasmid.
-I made 10 starters for In1 from the transformation plates- I picked them randomly.. 1 starter for T7*RNAP+pSB1AK3 (that I know it works) and 4 starters of native T3 RNAP+pSB1AK3 from the transformation plates (picked by random).
-Colony PCR for 2 colonies of In1 part, I stored them at 16C at the end of the PCR.
Asaf
I got clones from all the transformations including my negative control. I descided due to lack of time
to proside to the next PCR and in doing so I will make sure if I got the right insert transformed.
I made starters from different clones and also streak plates.