IGEM:Harvard/2007/Plates

Colony Count of Overnight Ligation vs. Shorter (2 hr Ligation)

Plated 6-20-07

Cell Type	Volume Plated	Colony Count (Short Ligation)	Colony Count (Overnight Ligation)
OmpA1 + random library, PCR	275 µL	5	41
OmpA1 + random library, PCR	25 µL	0	2
OmpA1 + random library, extension	275 µL	0	30
OmpA1 + random library, extension	25 µL	0	0
OmpA2 + random library, PCR	275 µL	0	1
OmpA2 + random library, PCR	25 µL	0	0
OmpA2 + random library, extension	275 µL	12	77
OmpA2 + random library, extension	25 µL	0	1

Colony Count of Overweekend Ligation

Plated 6/25/07 The following were all done with 275 uL:

Cell Type	Colony Count (Overweekend Ligation;1uL DNA)	Colony Count (Overweekend Ligation;6uL DNA)
OmpA1/Group1/S	0	127
OmpA1/Group1/S2	8	24
OmpA1/Group2/B	10	119
OmpA1/Group2/S2	4	79

Colony Count of Electroporation

Plated 7/11/07

Plate	Dilution of cells	0ng DNA	25ng DNA	50ng DNA	100ng DNA	250ng DNA	500ng DNA
LB-KAN	10^-2			521	136	197	361
LB	10^-3	489	329	576	267	130	338

MACS and FACS Cell Sort with AIDA Construct

Plated 7-20-07

Cell Type	Volume Plated	Colony Count (white)	Colony Count (red)
AIDA1 + strep, MACS	5 µL	5	0
AIDA1 + strep, MACS	50 µL	31	5
AIDA1 + strep, MACS	300 µL	235	10
AIDA1 + his, MACS	5 µL	0	1
AIDA1 + his, MACS	50 µL	18	0
AIDA1 + his, MACS	300 µL	206	11

Cell Type	Volume Plated	Colony Count (white)	Colony Count (red)
AIDA1 + strep, (+) fraction, FACS	10 µL	0	19
AIDA1 + strep, (-) fraction, FACS	10 µL	0	18
AIDA1 + his, (+) fraction, FACS	10 µL	187	16
AIDA1 + his, (-) fraction, FACS	10 µL	0	12 MACS Cell Sort with AIDA Construct-All Induced Plated 8-5-07 Cell Type Volume Plated Colony Count (white) Colony Count (red) AIDA1 + strep, (+) fraction, MACS 120 µL 101 3 AIDA1 + strep, (+) fraction, MACS 25 µL 13 2 AIDA1 + his, (+) fraction, MACS 120 µL 29 22 AIDA1 + his, (+) fraction, MACS 25 µL 5 4