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 |
