Team:UNICAMP-Brazil/Notebooks/September 6

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* Once we confirmed the isolation of both finO and finP sequences from the R plasmid, we now proceed to the purification of the PCR's product. It's quite important to purify the reaction since it will be used for digestion and later ligation.
* Once we confirmed the isolation of both finO and finP sequences from the R plasmid, we now proceed to the purification of the PCR's product. It's quite important to purify the reaction since it will be used for digestion and later ligation.
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*We purified both PCR's products by running an agarose gel of the entire product and then extracting the desirable band from the gel. We used Invitrogen's PureLink Quick Gel Extraction Kit to perform the purification, following the manufacturer's protocol without modifications.
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*We purified both PCR's products by running an agarose gel of the entire product and then extracting the desirable band from the gel. We used Invitrogen's PureLink Quick Gel Extraction Kit to perform the purification, following the manufacturer's protocol without modifications (Protocol 7, in Protocols section).
*After the procedure, we ran an agarose gel with the resulting product in order to confirm purification. We purified both finO and finP!
*After the procedure, we ran an agarose gel with the resulting product in order to confirm purification. We purified both finO and finP!

Revision as of 17:11, 16 September 2009

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YeastGuard

New biobricks

Today we digested all our 4 new biobrick parts with both enzymes: Xba1 and Spe1 (Promoter of JEN1 from K. lactis; Promoter of DLD1 from K. lactis; Lysozyme from G. gallus; Gene JEN1 from K. lactis). We also digested the biofusion vector with the same enzymes for future ligation.

GEL

  • Digestions that worked:

- Promoter of JEN1 from K. lactis

- Promoter of DLD1 from K. lactis

- Lysozyme from G. gallus

  • Digestions that didnt work:

- Gene JEN1 from K. lactis

- Biofusion vector

ColiGuard

finO and finP Purification

  • Once we confirmed the isolation of both finO and finP sequences from the R plasmid, we now proceed to the purification of the PCR's product. It's quite important to purify the reaction since it will be used for digestion and later ligation.
  • We purified both PCR's products by running an agarose gel of the entire product and then extracting the desirable band from the gel. We used Invitrogen's PureLink Quick Gel Extraction Kit to perform the purification, following the manufacturer's protocol without modifications (Protocol 7, in Protocols section).
  • After the procedure, we ran an agarose gel with the resulting product in order to confirm purification. We purified both finO and finP!

Marcelo





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