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Team:University of Ottawa/10 July 2008
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Djedrysiak (Talk | contribs) (New page: __TOC__ ==Today in the lab== '''Dan''' :'''Gel of S&D&T PCR products''' ::<li>The bands I needed were on the gel, they were somewhat close to non specific binding bands, so I took extra ca...) |
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'''Looking back''' | '''Looking back''' | ||
::<li> After Matt encountered problems with reduced enzymatic activity of restriction enzymes, I decided to redigest and reconfirm | ::<li> After Matt encountered problems with reduced enzymatic activity of restriction enzymes, I decided to redigest and reconfirm | ||
| + | '''Matt''' | ||
| + | :'''PCR confirmation''' | ||
| + | ::<li> Tested absorbance and Ran a PCR confirmation of IP producing cells (97 plasmid int. in BY4742 yeast strain) using both wild type BY4742 and H20 as control's. | ||
| + | :'''Digestion of PTP2 Ligation Product''' | ||
| + | ::<li> Another digestion of PTP2 was attempted using EcoRI - sadly I did not get the bands I needed, it seems the PTP2 was not correctly integrated into the Vector. | ||
Revision as of 18:50, 16 July 2008
Contents |
Today in the lab
Dan
- Gel of S&D&T PCR products
- The bands I needed were on the gel, they were somewhat close to non specific binding bands, so I took extra caution when cutting them out.
- After using the gel extraction kit the obtained concentrations were decent.
- PCR amplification of S&D&T
- Performed PCR on the S&D&T obtained products, if it works, It would be easy to obtain more DNA and would save the hassle of going back to step 1.
- Atcre and construct 1 concentrations
- Both these DNA fragments resulted in very low concentrations following gel extraction. Chris went ahead and used Atcre anyways. The concentration of construct 1 was too low to use.
- Talked to Cory and realized that AgeI could be used instead of ClaI. That would solve our problem of performing ligation on a 2bp overhang.
- We ordered new primers today for this.
Looking back
- After Matt encountered problems with reduced enzymatic activity of restriction enzymes, I decided to redigest and reconfirm
Matt
- PCR confirmation
- Tested absorbance and Ran a PCR confirmation of IP producing cells (97 plasmid int. in BY4742 yeast strain) using both wild type BY4742 and H20 as control's.
- Digestion of PTP2 Ligation Product
- Another digestion of PTP2 was attempted using EcoRI - sadly I did not get the bands I needed, it seems the PTP2 was not correctly integrated into the Vector.
