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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
|
| 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
|
MACS and FACS Cell Sort with AIDA Construct
Plated 7-20-07
| 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
|
