IGEM:Harvard/2007/Plates

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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

MACS Cell Sort with AIDA Sender Constructs

Plated 8-10-07

Note - Red colonies are what we want, white is background

Cell Type Volume Plated Colony Count
(red)
Colony Count
(white)
AIDA1 + strep sender, (+) fraction, MACS 75 µL 325 24
AIDA1 + strep sender, (+) fraction, MACS 20 µL 103 8
AIDA1 + his sender, (+) fraction, MACS 75 µL 588 49
AIDA1 + his sender, (+) fraction, MACS 20 µL 317 30