Team:UT-Dallas/Notebook/8-21

From 2014.igem.org

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<p class="tab"> Today is a medium day. </p>
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<p class="tab"> Today is a long day. </p>
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<p class="tab"> <b>Second batch</b>: We inoculated some toxT gRNA. Today we are minipreping it and test digest.</p>
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<p class="tab"> <b>Second batch</b>: We inoculated some toxT gRNA. Today we are minipreping and test digesting it.</p>
<p class="tab"> <b>Third batch (B3)</b>: Yesterday, we inoculated 2 reporter-Chlora colonies from each plates. Today we digest them, gel purify, ligate, and transform them onto Carb back bone, along with re-ligating and re-transforming ctxA that kept failing.</p>
<p class="tab"> <b>Third batch (B3)</b>: Yesterday, we inoculated 2 reporter-Chlora colonies from each plates. Today we digest them, gel purify, ligate, and transform them onto Carb back bone, along with re-ligating and re-transforming ctxA that kept failing.</p>
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<p class="tab"> <b>Forth batch (B4)</b>: We transformed B4 reporters on promoter-Chlora. </p>
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<p class="tab"> <b>Forth batch (B4)</b>: We transformed B4 reporters on promoter-Chlora. The plates are good today. There's one colonies on the negative and the colonies on other plates appear with different sizes, so we are picking 2 colonies from each plates to inoculate.</p>
<p>We remade some gRNA from the primers. Unfortunately, yesterday we didn't have the machine for overnight ligation. So we will ligate it tonight.</p>
<p>We remade some gRNA from the primers. Unfortunately, yesterday we didn't have the machine for overnight ligation. So we will ligate it tonight.</p>
<p class="tab"> Tomorrow: we should have colonies of B4 reporters on the plates we are transforming today.</p>
<p class="tab"> Tomorrow: we should have colonies of B4 reporters on the plates we are transforming today.</p>

Revision as of 17:04, 21 August 2014

Thursday, August 21, 2014

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Today is a long day.

Second batch: We inoculated some toxT gRNA. Today we are minipreping and test digesting it.

Third batch (B3): Yesterday, we inoculated 2 reporter-Chlora colonies from each plates. Today we digest them, gel purify, ligate, and transform them onto Carb back bone, along with re-ligating and re-transforming ctxA that kept failing.

Forth batch (B4): We transformed B4 reporters on promoter-Chlora. The plates are good today. There's one colonies on the negative and the colonies on other plates appear with different sizes, so we are picking 2 colonies from each plates to inoculate.

We remade some gRNA from the primers. Unfortunately, yesterday we didn't have the machine for overnight ligation. So we will ligate it tonight.

Tomorrow: we should have colonies of B4 reporters on the plates we are transforming today.

Today's tasks:

  • Miniprep ctxA, ctxB.
  • Test digest and ran gel for ctxA, ctxB.
  • Inoculate 2 colonies from each plate transformed yesterday (second batch, reporter vectors on Carb).
  • Prepared for sequencing: diluted to the correct volume and concentration.
  • Purified RBS (digested overnight yesterday).
  • Transformed of overnight ligation yesterday (third batch, reporter vectors with promoters under Chlora).
  • Inoculate more colonies from ctxA, ctxB plates.



RBS-Carb Gel was good.


Updating the wiki notebook from the slowest computer in the building because currently we can't log into any other computer. Rishika is gel-ing the ctx-s.