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Friday, July 29

Friday, 7/29

Members Present: Alex, Cathy, Zarifah, Celine, Hamed

LAB:

Morning:

●

Made 250ml of LB+agar+Cam and LB+agar+Kana and 500ml of LB liquid media

○

Disposed of the old LB liquid media (less than 150ml left in there)

○

Kayla has provided us a microcentrifuge tube of prepared Kanamycin which has been put in the pink Cam rack in the -20^oC freezer in WB403

●

LB+Agar+Kana was made using 1000x diluted Kanamycin that was supplied by Kayla

○

250ml LB+Agar with 250ul of Kanamycin stock

Afternoon:

●

Transformation on the ligations from the previous day:

○

Used CCD cells (50ul) with 1.5ul of the following plasmids:

I.

BB Digested

II.

BB Undigested

III.

BB DpnI

IV.

HISR+pCOLA

○

Also created two controls:

I.

Negative

●

50ul CCD

●

1.5ul NF water

II.

Blank

●

51.5ul of NF water

○

SOC pre-warmed at 37^oC, water bath at 42^oC

■

NOTE OF HUMAN ERROR

●

During heat shock, BB Digested and Negative floated away therefore exposed to water bath for an extra ~15 sec (for a total heat shock of ~60 sec)

●

750ul instead of 250ul of SOC was added to BB Undigested by accident

○

The following plates were made:

I.

BB Digested on LB+Cam (two duplicates: one fresh, one old)

II.

BB Undigested on LB+Cam (two duplicates: one fresh, one old)

III.

BB DpnI on LB+Cam (two duplicates: one fresh, one old)

IV.

Negative on LB+Cam (fresh plate)

V.

Blank/NF water on LB+Cam (fresh plate)

VI.

HISR+pCOLA on LB+Kana (two duplicates)

VII.

Negative/blank on LB+Kana (nothing streaked on, just a plate sitting in incubation)

○

All plates were stored inverted in the stationary incubator in WB403 at 37^oC

Administrative:

●

OG meeting with Alex, Ben and Anthony has been canceled and moved to Wednesday, August 3rd at 10am

●

Key VP21 is no longer able to open WB407, need to contact Susie

●

oGEM P&P Skype meeting:

○

Still no server, Fahim allocated tasks to each oGEM team to begin development of the Synthetic Biology Network Platform and they will be using local servers in the mean time

TO DO:

LAB TEAM:

●

Running low on CCD competent cells, need to make more

●

Miniprep and overnight culture hopefully grown cells

LAB MANAGERS:

●

Things to wash