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Team:SouthBend-Mishawaka-HS/Notebook
From 2011.igem.org
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May
May19
(1)Poured 12 LB AMP plates:(2mg of Ampicillin to 250ml of Agar)
(2)Obtained a vial of 2 microliters of TOP-10 electrocompetant E. coli cells (44-0003/793762)
(3)Added 50 microliters of pure water from the Gemini machine
(4)Added 10 ul of water to well 6E of plate 2, which contained part F2620.
(5)Added 2 ul of the diluted DNA in 50 ul of transformation solution (50mm CaCl2, pH 6.1 BIO-RAD Transformation Solution control # 31000008916 recieved 1-17-11 GT).
(6)Incubate on ice for 5 minutes
(7)Heat shocked at 40 degrees Celcius for 50 seconds
(8)Incubated at 4 degrees Celcius for 5 minutes
(9)Added 1mL of LB (5-17-11 DG)
(10)Plated 100 microliters and 25 microliters of transformed cells on LB-AMP (5-19-11 "crew")
May 18
We found no growth on the plates. We are going to re-transform them next week, this time using electricity instead of heat-shock.
May 24
(1)Obtained 40 microliters of electrocompetent TOP-10 E. coli cells.
(2)Added 2 microliters of F2620.
(3)Zapped with 1.8kV for 5.8ms.
(4)Recovered with 900 microliters of SOB to recover cells
(5)Incubated for 1hr. Then plated the newly transformed cells.
