Team:UNICAMP-Brazil/Notebooks/September 21

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(ColiGuard)
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*<p style=”text-align:justify;”>We run 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;”>We run 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>
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*<p style=”text-align:justify;”>Therefore, we will perform another miniprep of the cultures by which we best amplified our expected fragment.</p>
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*<p style=”text-align:justify;”>Therefore, we will perform another miniprep of the cultures by which we best amplified our expected fragment (highlighted).</p>
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[[image:PCRminipreps2309_bbfinO_bbfinP.jpg]]
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''Marcelo''
''Marcelo''
{{:Team:UNICAMP-Brazil/inc_rodape}}
{{:Team:UNICAMP-Brazil/inc_rodape}}

Revision as of 16:58, 18 October 2009

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ColiGuard

Digested F plasmid's electroelution

  • The digested F plasmid (digested with HindIII) was electroeluted, according to Protocol 12. A 110 V voltage was applied during 3h before its extraction from the membrane.

  • 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.

Gabriel

finO and finP's biobricks confirmation

  • We repeated today the PCR's of the minipreps samples, once we now have received new Taq Polymerase enzymes.

  • We run 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.

  • 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.

  • Therefore, we will perform another miniprep of the cultures by which we best amplified our expected fragment (highlighted).

PCRminipreps2309 bbfinO bbfinP.jpg


Marcelo