Team:UNICAMP-Brazil/Notebooks/September 12

From 2009.igem.org

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(PCR: Cre-Recombinase without ATG)
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*<p style=”text-align:justify;”>Since all of our fragments are correctly digested, we started today the ligation of both finO and finP sequences to the vector pSB1A3. Each sequence was ligated singly to the vector and, only after each one is confirmed, we'll join then both to a single construction.</p>
*<p style=”text-align:justify;”>Since all of our fragments are correctly digested, we started today the ligation of both finO and finP sequences to the vector pSB1A3. Each sequence was ligated singly to the vector and, only after each one is confirmed, we'll join then both to a single construction.</p>
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*<p style=”text-align:justify;”>We performed the ligation finO+vector pSB1A3 for an O/N period, according to protocol Y (see Protocols section).</p>
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*<p style=”text-align:justify;”>We performed the ligation finO+vector pSB1A3 for an O/N period, according to [https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/T4_DNA_Ligase Protocol 11].</p>
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''Marcelo''
====finP+pSB1A3 ligation====
====finP+pSB1A3 ligation====

Revision as of 15:32, 18 October 2009

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ColiGuard

finO+pSB1A3 ligation

  • Since all of our fragments are correctly digested, we started today the ligation of both finO and finP sequences to the vector pSB1A3. Each sequence was ligated singly to the vector and, only after each one is confirmed, we'll join then both to a single construction.

  • We performed the ligation finO+vector pSB1A3 for an O/N period, according to Protocol 11.

Marcelo

finP+pSB1A3 ligation

  • As we did for finO sequence, we also ligated finP to the vector pSB1A3 according to the same protocol. The ligation also lasted O/N period.

Marcelo

PCR: Cre-Recombinase without ATG

  • Today we realized again the PCR reaction to isolate the Cre-Recombinase without the ATG codon. The amplification was succesfully realized, being comproved with an 1% agarose gel run. The photo proves the amplification and the expected size of our fragment:

YeastGuard

New biobricks in biobrick format

  • E. coli transformed with the new biobricks grew very well on the LB+AMP plates!

Taís