Team:UNICAMP-Brazil/Notebooks/September 21

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(New page: ====Digested F plasmid's electroelution==== *The digested F plasmid (digested with HindIII) was electroeluted, according to protocol 9. A 110 V voltage was applied during 3h before its ex...)
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__NOTOC__
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==''' ColiGuard '''==
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====Digested F plasmid's electroelution====
====Digested F plasmid's electroelution====
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*The digested F plasmid (digested with HindIII) was electroeluted, according to protocol 9. A 110 V voltage was applied during 3h before its extraction from the membrane.
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*<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>
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*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.
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*<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>
''Gabriel''
''Gabriel''
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====finO and finP's biobricks confirmation====
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*<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>
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*<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>
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*<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 (highlighted).</p>
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[[image:PCRminipreps2309_bbfinO_bbfinP.jpg]]
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''Marcelo''
{{:Team:UNICAMP-Brazil/inc_rodape}}
{{:Team:UNICAMP-Brazil/inc_rodape}}

Latest revision as of 02:57, 22 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 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.

  • 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




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