Team:UNICAMP-Brazil/Notebooks/September 13

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====Purification: Cre-Recombinase without ATG====
====Purification: Cre-Recombinase without ATG====
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*<p style=”text-align:justify;”>Today we realized the purification of the Cre-Recombinase without the ATG codon that was amplified september 12.
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*<p style=”text-align:justify;”>Today we realized the purification of the Cre-Recombinase without the ATG codon that was amplified september 12.</p>
*<p style=”text-align:justify;”>We purified the PCR's product 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 ([https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Purification_of_DNA_fragments_from_agarose_gels Protocol 7]).</p>
*<p style=”text-align:justify;”>We purified the PCR's product 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 ([https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Purification_of_DNA_fragments_from_agarose_gels Protocol 7]).</p>
*<p style=”text-align:justify;”>After the procedure, we ran an agarose gel with the resulting product in order to confirm purification. We purified the Cre-Recombinase without ATG codon!</p>
*<p style=”text-align:justify;”>After the procedure, we ran an agarose gel with the resulting product in order to confirm purification. We purified the Cre-Recombinase without ATG codon!</p>

Revision as of 20:32, 17 October 2009

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ColiGuard

finO+pSB1A3

  • After the O/N period, today we transformed the ligation finO+pSB1A3 into electrocompetent E. coli bacteria, strain DH10B. We followed Protocol 3, without modifications.

  • We then plated the transfomed cells in LB-AMP medium, and let them grow at 37ºC for an O/N period.

finP+pSB1A3

  • We followed the same produce for the finP+pSB1A3 ligation.

Marcelo

Purification: Cre-Recombinase without ATG

  • Today we realized the purification of the Cre-Recombinase without the ATG codon that was amplified september 12.

  • We purified the PCR's product 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).

  • After the procedure, we ran an agarose gel with the resulting product in order to confirm purification. We purified the Cre-Recombinase without ATG codon!

Víctor

YeastGuard

New biobrick - The first screening

  • 6 colonies of each new biobrick were chosen of the plates made yesterday to grow in liquid LB.

  • Then, we performed miniprep to get our new biobricks purified (Protocol 2).

  • We then digested the plasmids obtained by the miniprep with XbaI to confirm the correct ligation of the parts following the biobrick format (Protocol 11). However, the electrophoresis gel (shown below) showed an unexpected pattern of bands that can't confirm correctly ligations of our biobricks. =(

‎

Wesley and Gleidson




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