"
Team:UNICAMP-Brazil/Notebooks/September 21
From 2009.igem.org
(→ColiGuard) |
(→finO and finP's biobricks confirmation) |
||
| (2 intermediate revisions not shown) | |||
| Line 7: | Line 7: | ||
====Digested F plasmid's electroelution==== | ====Digested F plasmid's electroelution==== | ||
| - | *<p style=”text-align:justify;”>The digested F plasmid (digested with '' | + | *<p style=”text-align:justify;”>The digested F plasmid (digested with ''Hind''III) was electroeluted, according to [https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Electroelution Protocol 12]. A 110 V voltage was applied during 3h before its extraction from the membrane.</p> |
*<p style=”text-align:justify;”>The next step is to verify if the electroelution went OK. The collected samples of purified plasmid will be applied to a 0,8% agarose gel.</p> | *<p style=”text-align:justify;”>The next step is to verify if the electroelution went OK. The collected samples of purified plasmid will be applied to a 0,8% agarose gel.</p> | ||
| Line 15: | Line 15: | ||
*<p style=”text-align:justify;”>We repeated today the PCR's of the minipreps samples, once we now have received new Taq Polymerase enzymes.</p> | *<p style=”text-align:justify;”>We repeated today the PCR's of the minipreps samples, once we now have received new Taq Polymerase enzymes.</p> | ||
| - | *<p style=”text-align:justify;”>We | + | *<p style=”text-align:justify;”>We ran an agarose gel of the reaction products and, according to it, we amplified in a few samples a fragment with the expected size of finO! We couldn't verify this amplification for finP though.</p> |
*<p style=”text-align:justify;”>Nevertheless, we also got several inespecific amplification in almost all samples. We considered that this was caused by the increased amount of genomic DNA we obtained after the miniprep procedure.</p> | *<p style=”text-align:justify;”>Nevertheless, we also got several inespecific amplification in almost all samples. We considered that this was caused by the increased amount of genomic DNA we obtained after the miniprep procedure.</p> | ||
| - | *<p style=”text-align:justify;”>Therefore, we will perform another miniprep of the cultures by which we best amplified our expected fragment.</p> | + | *<p style=”text-align:justify;”>Therefore, we will perform another miniprep of the cultures by which we best amplified our expected fragment (highlighted).</p> |
| + | |||
| + | [[image:PCRminipreps2309_bbfinO_bbfinP.jpg]] | ||
| + | |||
''Marcelo'' | ''Marcelo'' | ||
{{:Team:UNICAMP-Brazil/inc_rodape}} | {{:Team:UNICAMP-Brazil/inc_rodape}} | ||
Latest revision as of 02:57, 22 October 2009
| ||||||||||||||||||||||||||||||||||
