Team:UNICAMP-Brazil/Notebooks/September 18

(Difference between revisions)

Revision as of 00:30, 20 October 2009

Today we performed mini-preps (Protocol 2) to extract the plasmids from the selected colonies inoculated yesterday.

After the extraction we performed PCR reactions with 3 pairs of primers to confirm the ligation:

- Ppy-F and Ppy-R

- Ppy-F and VR

- VF and VR

Unfortunately, the sizes of the bands in the agarose gel were not compatible with the expected size of the bands. =(

We tried to find the colonies containing the biobricks by colony PCR (using the forward primer of the insert and the reverse of the plasmid). We did 10 reactions of each biobrick. Unfortunately we didn´t find the correct bands.

GEL

Taís

We decided to make a bigger screening of our colonies to find the ones containing the biobricks (pJen1, pDLD, Lysozyme, JEN1 orf). We grew 24 colonies from each biobrick O.N. in liquid medium.

Raíssa

@@ Line 2: / Line 2: @@
 {{:Team:UNICAMP-Brazil/inc calendar}}
 __NOTOC__
+==''' ColiGuard '''==
+==== PY Promoter - Mini-prep and PCR confirmation ====
+*<p style=”text-align:justify;”>Today we performed mini-preps ([https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Mini-Prep Protocol 2]) to extract the plasmids from the selected colonies inoculated yesterday.</p>
+*<p style=”text-align:justify;”>After the extraction we performed PCR reactions with 3 pairs of primers to confirm the ligation:</p>
+<p style=”text-align:justify;”>- Ppy-F and Ppy-R</p>
+<p style=”text-align:justify;”>- Ppy-F and VR</p>
+<p style=”text-align:justify;”>- VF and VR</p>
+*<p style=”text-align:justify;”>Unfortunately, the sizes of the bands in the agarose gel were not compatible with the expected size of the bands. =(</p>
 ==''' YeastGuard '''==