Today is a long day.

Second batch: We inoculated some toxT gRNA. Today we are minipreping it. (It already passed the test digest.

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 B3 reporters on the plates we are transforming today. We should miniprep the B4 reporters we inoculated today. For gRNA of B4, ligation will be done and we will transform them tomorrow. Saturday, we will inoculate. Sunday, we will miniprep. Next week, on Monday, we should have everything ready for test digest and then finally send the last batch of sequencing!

• 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.