Team:UNICAMP-Brazil/Notebooks/September 6

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==''' ColiGuard '''==
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====finO and finP Purification====
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*<p style=”text-align:justify;”>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.</p>
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*<p style=”text-align:justify;”>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 ([https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Purification_of_DNA_fragments_from_agarose_gels Protocol 7]).</p>
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*<p style=”text-align:justify;”>After the procedure, we ran an agarose gel with the resulting product in order to confirm purification. We purified both finO and finP!</p>
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''Marcelo'' 
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==== Cre Recombinase ====
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*<p style=”text-align:justify;”>Today we performed minipreps of Cre-Recombinase´s biobrick, that we confirmed by running an electrophoresis agarose gel.</p>
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''Víctor''
==''' YeastGuard '''==
==''' YeastGuard '''==
====New biobricks====
====New biobricks====
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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.
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*<p style=”text-align:justify;”>Today we digested all our 4 new biobrick parts with both enzymes: ''Xba''I and ''Spe''I (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.</p>
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GEL
 
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* Digestions that worked:
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[[Image:20090905_PCR_partes_BioF.png|300px|center|‎]]
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- Promoter of JEN1 from K. lactis
 
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- Promoter of DLD1 from K. lactis
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* Digestions that worked:
-
- Lysozyme from G. gallus
+
- Promoter of JEN1 from ''K. lactis''
-
* Digestions that didnt work:
+
- Promoter of DLD1 from ''K. lactis''
-
 
+
-
- Gene JEN1 from K. lactis
+
- Biofusion vector
- Biofusion vector
-
==''' ColiGuard '''==
 
 +
* Digestions that didnt work:
-
====finO and finP Purification====
+
- Gene JEN1 from ''K. lactis''
-
* 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.
+
- Lysozyme from ''G. gallus''
-
*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.
+
-
*After the procedure, we ran an agarose gel with the resulting product in order to confirm purification. We purified both finO and finP!
+
-
''Marcelo'' 
 
{{:Team:UNICAMP-Brazil/inc_rodape}}
{{:Team:UNICAMP-Brazil/inc_rodape}}

Latest revision as of 02:12, 22 October 2009

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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).

  • After the procedure, we ran an agarose gel with the resulting product in order to confirm purification. We purified both finO and finP!

Marcelo

Cre Recombinase

  • Today we performed minipreps of Cre-Recombinase´s biobrick, that we confirmed by running an electrophoresis agarose gel.

Víctor

YeastGuard

New biobricks

  • Today we digested all our 4 new biobrick parts with both enzymes: XbaI and SpeI (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.


‎


  • Digestions that worked:

- Promoter of JEN1 from K. lactis

- Promoter of DLD1 from K. lactis

- Biofusion vector


  • Digestions that didnt work:

- Gene JEN1 from K. lactis

- Lysozyme from G. gallus





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