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Friday June 17, 2016

Friday, 6/17

Members Present: Hamed, Kat, Marc, Dk

LAB:

Morning:

- Executed the S12 protocol; cells are forzen at -80C and are ready for sonication (buffer B has also been prepared)

- Asked Kayla for a new protocol for making chemically competent cells. We have now ordered the materials required for this.

- Results of the second attempt for using competent cells: Note that CCA is the first batch of chemically competent cells, CCB is the second batch of chemically competent cells, and CC0 refers to the chemically competent cells obtained from Kayla. Also note that CCA and CCB were rendered competent through Magnesium chloride and CC0 through rubidium chloride.

3X comp - 0.5pg.jpg

0.5 pg plated on both CCA and CCB cells. Note that no colonies grew.

3X comp - 10pg.jpg

CCA and CCB cells with 10pg. Note that CCA had 5 colonies and CCB had 4 colonies.

3X comp - 50pg.jpg

50pg of RFP plated on both CCA, CCB and CCo. Note that CCA had 16 colonies, CCB had 6 colonies, and CC0 had 369 colonies. This demonstrates that the CC0 cells obtained from Kayla have a much higher transformation efficiency.

3X comp - RFP.jpg

Miniprepped RFP 1B transformed with CCA, CCB and CC0 cells. 478 colonies on CCA, 565 colonies, and overgrowth following transformation with CC0 cells.

3X - negative control.jpg

Negative control with CCA and CCB, wherein no plasmid was added during transformation.

3X.jpg

Streaked some RFP expressing bacterial on LB + CAM plates. We were initially worried that our CAM concentration might be too high, so this was to verify that the cells containing the resistance gene could survive.

Afternoon:

- Sonication and Aliquoting S12 mix (stored at -80C); RFP plasmid was tested in the extract (results to be recorded on Monday)

Administrative:

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First aid kit + waterproof bandages + hydrogel can be found in the communal locker

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Liquid nitrogen bucket can be found in 303 (we can use this with permission)

TO DO:

For the next day:

LAB TEAM:

LAB MANAGERS:

Gmail correspondence:

Meetings/Notes:

References: