pGBR22

The pGBR22 DNA was eluted today using a high speed midi kit and stored in 2 tubes in 50ul aliquots. One tube is in the top loading freezer in Dr. B’s box and the other in the mini VDS fridge. The concentration of the DNA read at absorbance 260nm was taken twice : 161.1ng/ul and 165.5ng/ul.

PTP1B Expression

I have added IPTG to the 2 2L flasks in the shaker around 11:30 and they need to shake for about 4 hours. Be careful about the fat flask, I jimmy-rigged it to fit in the shaker, but I would keep an eye on it just to make sure it does not spill over. Also, there is orange tape on the flasks with all the steps I have done this morning.

Can one of the mentors spin down today and store in -20 C fridge, and take a sample (sample 1) of the culture after the four hr induction period?
Also, if someone wouldn’t mind drying the gels above the VDS cubby hole, that would be great. There is cellophane in the drawer where the drying supplies are. If someone can’t get to them, that is fine. The PTP1B expression is priority.

Thanks,
Leighanna

LB Media

I could only find 2 2L flasks in the painter 2.14 lab. I have both flasks and 1L of LB autoclaving right now. I will also run another PAGE gel of Thao’s and Yiling’s elution samples today to determine if an FPLC can be done on PTP1B elutions.

Imaging PAGE gels

Dr. B, I have uploaded the gels onto the google docs under the mentor’s folder. Thao’s gel didn’t look too good, not even the marker was really visible. I think it may have just been the way the gel ran, but Yiling’s gel as well as my gel for InhA turned out looking okay. I will dry them on Monday.

PAGE Gels staining

Dr. B, the PAGE gels are in the shaker staining as of 2:45pm today.
If you would please, destain overnight, I will come into tomorrow to take a quick image of them and either store it on google docs or email you the pictures.

Thanks,
Leighanna

Nanodrop and gels

The nanodrop is working and well on the computer across from where it used to sit.
In regards to the PAGE gels, I was just wondering how many everyone needed. I was planning to make about 5. Mentors let me know if I need to make more or less. Thank you.

Radhika’s plates are now in the 4 degree VDS fridge

It’s done.