Team:UNICAMP-Brazil/Notebooks/October 16

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*<p style=”text-align:justify;”>To perform our tests, we digested the following minipreps: pJEN1+YFP, pJEN1+Lys, pDLD+YFP, pDLD+Lys with ''Xba''I and ''Pst''I to connect to YEP vector. We also digested the plasmid YEP+Adh1-YFP to confirm the correct insertion of the part into YEP vector. The digestion showed that our new devices are ricght! =) But there is something wrong with the YEP+Adh1-YFP construction, it seams the digestion were incomplete.</p>
*<p style=”text-align:justify;”>To perform our tests, we digested the following minipreps: pJEN1+YFP, pJEN1+Lys, pDLD+YFP, pDLD+Lys with ''Xba''I and ''Pst''I to connect to YEP vector. We also digested the plasmid YEP+Adh1-YFP to confirm the correct insertion of the part into YEP vector. The digestion showed that our new devices are ricght! =) But there is something wrong with the YEP+Adh1-YFP construction, it seams the digestion were incomplete.</p>
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GEL
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[[Image:digested-devices.jpg|600px|center]]
*<p style=”text-align:justify;”>We purified one digested fragment of each device: pJEN1+YFP (1781bp), pJEN1+Lys(1567bp), pDLD+YFP (1246bp), pDLD+Lys (1032bp). We ligated the digested fragments with YEP vector and transformed in competent ''E. coli''.</p>
*<p style=”text-align:justify;”>We purified one digested fragment of each device: pJEN1+YFP (1781bp), pJEN1+Lys(1567bp), pDLD+YFP (1246bp), pDLD+Lys (1032bp). We ligated the digested fragments with YEP vector and transformed in competent ''E. coli''.</p>
*<p style=”text-align:justify;”>We did miniprep of YEP+Adh1-lysozyme. Later we digested the plasmids to confirm the insertion of the Adh1-Lysozyme construction into YEP. We also digested 10 more plasmids YEP-Adh1-YFP again, this time we digested a little longer to try to avoid the incomplete digestion problem.</p>
*<p style=”text-align:justify;”>We did miniprep of YEP+Adh1-lysozyme. Later we digested the plasmids to confirm the insertion of the Adh1-Lysozyme construction into YEP. We also digested 10 more plasmids YEP-Adh1-YFP again, this time we digested a little longer to try to avoid the incomplete digestion problem.</p>
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GEL
+
[[Image:miniprepYEP+.jpg|400px|center]]
*<p style=”text-align:justify;”>We transformed competent yeasts with YEP+Adh1-YFP without confirming the correct ligation.</p>
*<p style=”text-align:justify;”>We transformed competent yeasts with YEP+Adh1-YFP without confirming the correct ligation.</p>

Revision as of 05:06, 21 October 2009

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YeastGuard

Yeast experiments

  • To perform our tests, we digested the following minipreps: pJEN1+YFP, pJEN1+Lys, pDLD+YFP, pDLD+Lys with XbaI and PstI to connect to YEP vector. We also digested the plasmid YEP+Adh1-YFP to confirm the correct insertion of the part into YEP vector. The digestion showed that our new devices are ricght! =) But there is something wrong with the YEP+Adh1-YFP construction, it seams the digestion were incomplete.

Digested-devices.jpg

  • We purified one digested fragment of each device: pJEN1+YFP (1781bp), pJEN1+Lys(1567bp), pDLD+YFP (1246bp), pDLD+Lys (1032bp). We ligated the digested fragments with YEP vector and transformed in competent E. coli.

  • We did miniprep of YEP+Adh1-lysozyme. Later we digested the plasmids to confirm the insertion of the Adh1-Lysozyme construction into YEP. We also digested 10 more plasmids YEP-Adh1-YFP again, this time we digested a little longer to try to avoid the incomplete digestion problem.

MiniprepYEP+.jpg

  • We transformed competent yeasts with YEP+Adh1-YFP without confirming the correct ligation.

  • We did the YNP Ura- solid medium.

Raíssa and Taís





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