SBB12Ntbk-Hyungjun Kim

Hyungjun Kim March 15th

-I performed a digest on the second set of mini-prepped sbb1203 and sbb1224 parts.

-The analytic digests look good! Sent in for sequencing.

Hyungjun Kim March 13th

-The sbb1224 from my first set of products are duds. Nothing showed up even on the second analytical mapping.

-For my second set of products, the colonies Zach picked from sbb1224 and sbb1203 seemed to have grown fine. I mini prepped them, asnd hopefully these products will show up on a analytical digest.

Hyungjun Kim March 8th

-I went ahead digested the mini-prepped sbb1224 product from last time. I ran it on a gel. Got some DNA this time around but size was way too big. I think it's worth doing a redigest, I may have messed up the digest.

-I also did a ligation reaction on the 2nd PCA amp product I saved previously and transformed, and plated. Hopefully these work well.

-For my extreme back-up run, I set up the 2nd PCA amp step.

Hyungjun Kim March 6th

-All the colonies I picked for sbb1203 didn't grow...big surprise.

-Zach picked 2 sbb1124 colonies for me and they grew. I finished the miniprep on that. I really hope these don't run blank on gels like last time.

-I also took my products from the PCA amp step for sbb1224 and sbb1203 and digested them. I'm starting over from this step.

-I also started a parallel run from the first PCA step just in case.

-Finished small frag zymo on sbb1203 and zymo gel purification on sbb1224.

Hyungjun Kim March 2st

-The plate for sbb1203 actually looked like E.Coli so that's a good sign. Went ahead and picked colonies. Keeping fingers crossed on this.

-I did a ligation and transformation on the sbb1224 digest part and the bca1834 vector digest.

-Plated the sbb 1224 transformation product, hopefully it's better or I'll have to go back a step further.

Hyungjun Kim March 1st

-None of the colonies grew for sbb 1203, so I went ahead to do another transformation and plated.

-I digested the sbb1224 minipreps and ran them on a analytical gel

-Well that was depressing, nothing came out on the DNA gel for sbb1224, so I guess I'll backtrack to the digested 1224 part and do a digestion/transformation again.

Hyungjun Kim February 28th

-Picked up the cell cultures that had been picked over the weekend by Zach

-The 3 tubes for sbb1224 looked good for mini preps while one dubious tube looked okay for sbb1203

-I went ahead and mini-prepped the 4 tubes.

-I also picked 4 more colonies for sbb 1203, hopefully these turn out fine

Hyungjun Kim February 24th

-Ran zymo cleanup on for the sbb1224 gel purification -Performed transformation with sbb1224 and sbb1224 into competent cells

Hyungjun Kim February 23th

-Ran zymo cleanups on the PCA amplification products -Did a NheI/BamI digest -I proceeded with a zymo on sbb1203 and did a gel purification on sbb 1224

Hyungjun Kim February 21th

-I ran zymo cleanups on the PCA products for sbb 1203 and 1224. -Small fragment cleanup was used for sbb1203 and regular zymo cleanup for 1224. -I set up the 2nd PCA amplification step for both parts.

Hyungjun Kim February 16th

-Started on making sbb1203 and sbb1224 parts -Ran the first PCA steps for sbb1203 and sbb1224

Hyungjun Kim

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