SBB12Ntbk-Hyungjun Kim

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Hyungjun Kim April 12th

Unfortunately the plates did not have many colonies grow. Our group is going to transform again doubling concentrations, and hopefully this will result in usable plates. 

Hyungjun Kim April 10th

Our analysis group went ahead and transformed the necessary plasmids into competent cells.(protocol is attached)

Hyungjun Kim April 5th

Asked Zach for necessary parts. They should be ready by next Tuesday. Planned out timeline for this experiment and coordinated with another group to share a plate for the plate reader.

Hyungjun Kim April 3rd

Planned for analysis experiments of sbb1223 camelid VHH heavy chain. We are going to characterize it's activity by quantitatively observing GFP expression.

Hyungjun Kim March 22nd

Anaylyzed DNA reads in a group again.

Hyungjun Kim March 20th

Anaylyzed DNA reads in a group.

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.

Image:NEB_2-log_ladder.gifImage:2012_03_15_gel2_ssb2012spring.jpg

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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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