Team:UNICAMP-Brazil/Notebooks/August 28

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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.

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

@@ Line 6: / Line 6: @@
 ==''' YeastGuard '''==
-====RBS+CFP====
-We did the permanent colonies.
-''Taís''
 ====''Kluyveromices lactis'' genes====
-The PCR for the ''K. lactis'' genes worked very well! =]  We got specific bands for the 3 sequences.
+*<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>
-GEL PCR
+====Terminator biobrick====
-''Taís''
+*<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.
-====Colony PCR – Terminator biobrick====
+[[Image:PCRKL.jpg‎|350px|center]]
-The colony PCR was made to confirm the presence of terminator transformants, because  the antibiotic resistence is not working to any antibiotics.
-We plated the the cells in LB, and picked some colonies with a toothpick, inoculate on a backup LB plate and put the toothpick on a tube with 20µl H2O, then, denature 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).
+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.
-We also innoculated ten colonies transformed with plasmids containing the terminator biobrick to do the miniprep tomorrow.
+*<p style=”text-align:justify;”>Preparing for Miniprep</p>
+We also inoculated 10 colonies transformed with the terminator biobrick to perform the miniprep tomorrow.
 ''Wesley''
 {{:Team:UNICAMP-Brazil/inc_rodape}}