Team:UNICAMP-Brazil/Notebooks/September 24

From 2009.igem.org

(Difference between revisions)
(→var page = wgPageName.substr(30);document.write(page.replace(/_/, " "));)
(→CeaB and CeiB: Miniprep and transformation)
 
(17 intermediate revisions not shown)
Line 1: Line 1:
-
==<html><script>var page = wgPageName.substr(30);document.write(page.replace(/_/, " "));</script></html>==
+
{{:Team:UNICAMP-Brazil/inc_topo}}
 +
{{:Team:UNICAMP-Brazil/inc calendar}}
 +
 
 +
__NOTOC__
==''' ColiGuard '''==
==''' ColiGuard '''==
Line 5: Line 8:
====''E. coli'' transformation with F plasmid====
====''E. coli'' transformation with F plasmid====
-
*The ligation reaction was assembled according to protocol 10. Both the plasmid samples were used to recircularization. The reaction tubes was kept O/N at 4°C.
+
*<p style=”text-align:justify;”>''E. coli'' DH10B electrocompetent cells were transformed using the recircularized F plasmid, according to [https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Electroporation Protocol 3].</p>
 +
*<p style=”text-align:justify;”>After the incubation time, 100 μl of medium containing the transformed cells were plated in LB-AMP-Xgal culture plates, and incubated O/N at 37°C.</p>
''Gabriel''
''Gabriel''
 +
 +
====finO and finP - Still Trying to Confirm our Biobricks====
 +
 +
*<p style=”text-align:justify;”>After the miniprep procedure, we performed PCRs with finO and finP's designed primer, in order to confirm that our inserted fragment is indeed contained in the vector.</p>
 +
*<p style=”text-align:justify;”>As we had already got an amplified fragment of expected size from those colonies, we expect that those PCRs results in a successful amplification of our fragments. Moreover, as we aimed in reducing the amount of genomic DNA, we should got less inespecific amplifications.</p>
 +
 +
 +
''Marcelo''
 +
 +
 +
 +
==== CeaB and CeiB: Miniprep and transformation ====
 +
 +
*<p style=”text-align:justify;”>Today, we performed miniprep ([https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Mini-prep Protocol 2]) of the transformed cells and PCR again, so that avoids unspecific amplification. We didn’t get the expected band size again.</p>
 +
 +
 +
''Luige''
{{:Team:UNICAMP-Brazil/inc_rodape}}
{{:Team:UNICAMP-Brazil/inc_rodape}}

Latest revision as of 03:10, 22 October 2009

Topo l2.gif topo_r_igem.gif
topo_r_b.gif
Back to Calendar

ColiGuard

E. coli transformation with F plasmid

  • E. coli DH10B electrocompetent cells were transformed using the recircularized F plasmid, according to Protocol 3.

  • After the incubation time, 100 μl of medium containing the transformed cells were plated in LB-AMP-Xgal culture plates, and incubated O/N at 37°C.

Gabriel

finO and finP - Still Trying to Confirm our Biobricks

  • After the miniprep procedure, we performed PCRs with finO and finP's designed primer, in order to confirm that our inserted fragment is indeed contained in the vector.

  • As we had already got an amplified fragment of expected size from those colonies, we expect that those PCRs results in a successful amplification of our fragments. Moreover, as we aimed in reducing the amount of genomic DNA, we should got less inespecific amplifications.


Marcelo


CeaB and CeiB: Miniprep and transformation

  • Today, we performed miniprep (Protocol 2) of the transformed cells and PCR again, so that avoids unspecific amplification. We didn’t get the expected band size again.


Luige




Retrieved from "http://2009.igem.org/Team:UNICAMP-Brazil/Notebooks/September_24"