Team:UNICAMP-Brazil/Notebooks/September 18
From 2009.igem.org
(→CeaB, CeiB and BBa_B0015 restriction, purification and quantification) |
(→New biobricks - Second screening) |
||
(3 intermediate revisions not shown) | |||
Line 27: | Line 27: | ||
====Cre-Recombinase and pSB1A3 digestion==== | ====Cre-Recombinase and pSB1A3 digestion==== | ||
- | *<p style=”text-align:justify;”>Today we digested the purified PCR product for Cre-Recombinase without ATG start codon (see Septermber 13th) with | + | *<p style=”text-align:justify;”>Today we digested the purified PCR product for Cre-Recombinase without ATG start codon (see Septermber 13th) with ''Xba''I and ''Spe''I restriction enzymes. We also digested the vector pSB1A3 with the same enzymes, aiming in the future ligation between them.</p> |
- | *<p style=”text-align:justify;”>Digestion lasted 3 hours on both cases. | + | *<p style=”text-align:justify;”>Digestion lasted 3 hours on both cases.</p> |
''Víctor'' | ''Víctor'' | ||
Line 35: | Line 35: | ||
====New biobricks - Second screening==== | ====New biobricks - Second screening==== | ||
- | This time 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), not digestion. We did 10 reactions of each biobrick. Unfortunately we didn´t find the correct bands.</p> | + | *<p style=”text-align:justify;”>This time 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), not digestion. We did 10 reactions of each biobrick. Unfortunately we didn´t find the correct bands.</p> |
[[image:20090918_PCRcol_todos_JEN1orf.JPG.png|350px|center]] | [[image:20090918_PCRcol_todos_JEN1orf.JPG.png|350px|center]] | ||
Line 42: | Line 42: | ||
''Taís'' | ''Taís'' | ||
- | *<p style=”text-align:justify;”>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.</p> | + | *<p style=”text-align:justify;”>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.</p> |
''Raíssa'' | ''Raíssa'' | ||
{{:Team:UNICAMP-Brazil/inc_rodape}} | {{:Team:UNICAMP-Brazil/inc_rodape}} |
Latest revision as of 03:42, 22 October 2009
|