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.

• 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.