Team:UNICAMP-Brazil/Notebooks/August 28

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====''Kluyveromices lactis'' genes====
====''Kluyveromices lactis'' genes====
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The PCR for the ''K. lactis'' genes worked very well! =]  We got specific bands for the 3 sequences.
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*<p style=”text-align:justify;”>The PCR for the ''K. lactis'' genes worked very well! =]  We got specific bands for the 3 sequences (agarose gel above).</p>
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GEL PCR 
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''Taís''
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====Terminator biobrick====
====Terminator biobrick====
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* Colony PCR
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*<p style=”text-align:justify;”>Colony PCR</p>
The colony PCR was made to confirm the presence of terminator transformants, because  the antibiotic resistence is not working to any antibiotics.  
The colony PCR was made to confirm the presence of terminator transformants, because  the antibiotic resistence is not working to any antibiotics.  
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We plated the cells in LB and picked up some colonies with a toothpick, inoculated on a backup LB plate and put the toothpick on a tube with 20µl H2O, then, denatured this cells 95°C, 5 minutes and heat shock on ice 5 minutes. Spin the tubes and use 1µl to make the PCR (see protocol).
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[[Image:PCRKL.jpg‎|350px|center]]
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We plated the cells in LB and picked up some colonies with a toothpick, inoculated on a backup LB plate and put the toothpick on a tube with 20µl H2O. Then, denatured these cells at 95°C during 5 minutes and heat shocked on ice for 5 minutes. Spinned the tubes and used 1 µl to make the PCR.
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* Preparing for Miniprep
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*<p style=”text-align:justify;”>Preparing for Miniprep</p>
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We also inoculated ten colonies transformed with the terminator biobrick to do the miniprep tomorrow.
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We also inoculated 10 colonies transformed with the terminator biobrick to perform the miniprep tomorrow.
''Wesley''
''Wesley''
{{:Team:UNICAMP-Brazil/inc_rodape}}
{{:Team:UNICAMP-Brazil/inc_rodape}}

Latest revision as of 01:50, 22 October 2009

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YeastGuard

Kluyveromices lactis genes

  • The PCR for the K. lactis genes worked very well! =] We got specific bands for the 3 sequences (agarose gel above).

Terminator biobrick

  • Colony PCR

The colony PCR was made to confirm the presence of terminator transformants, because the antibiotic resistence is not working to any antibiotics.

PCRKL.jpg


We plated the cells in LB and picked up some colonies with a toothpick, inoculated on a backup LB plate and put the toothpick on a tube with 20µl H2O. Then, denatured these cells at 95°C during 5 minutes and heat shocked on ice for 5 minutes. Spinned the tubes and used 1 µl to make the PCR.

  • Preparing for Miniprep

We also inoculated 10 colonies transformed with the terminator biobrick to perform the miniprep tomorrow.

Wesley




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