IGEM:Harvard/2007/Plates: Difference between revisions
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| OmpA1/Group2/S2||4||79 | | OmpA1/Group2/S2||4||79 | ||
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== Colony Count of Electroporation == | |||
Plated 7/11/07 | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''Plate''' | |||
| align="center" style="background:#f0f0f0;"|'''Dilution of cells''' | |||
| align="center" style="background:#f0f0f0;"|'''0ng DNA''' | |||
| align="center" style="background:#f0f0f0;"|'''25ng DNA''' | |||
| align="center" style="background:#ff0000;"|'''50ng DNA''' | |||
| align="center" style="background:#f0f0f0;"|'''100ng DNA''' | |||
| align="center" style="background:#f0f0f0;"|'''250ng DNA''' | |||
| align="center" style="background:#f0f0f0;"|'''500ng DNA''' | |||
|- | |||
| LB-KAN||10^-2|| || ||521||136||197||361 | |||
|- | |||
| LB||10^-3||489||329||576||267||130||338 | |||
|} | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''Cell Type''' | |||
| align="center" style="background:#f0f0f0;"|'''Volume Plated''' | |||
| align="center" style="background:#f0f0f0;"|'''Colony Count <br>(white)''' | |||
| align="center" style="background:#f0f0f0;"|'''Colony Count <br>(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 | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''Cell Type''' | |||
| align="center" style="background:#f0f0f0;"|'''Volume Plated''' | |||
| align="center" style="background:#f0f0f0;"|'''Colony Count <br>(white)''' | |||
| align="center" style="background:#f0f0f0;"|'''Colony Count <br>(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 | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''Cell Type''' | |||
| align="center" style="background:#f0f0f0;"|'''Volume Plated''' | |||
| align="center" style="background:#f0f0f0;"|'''Colony Count <br>(white)''' | |||
| align="center" style="background:#f0f0f0;"|'''Colony Count <br>(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 | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''Cell Type''' | |||
| align="center" style="background:#f0f0f0;"|'''Volume Plated''' | |||
| align="center" style="background:#f0f0f0;"|'''Colony Count <br>(red)''' | |||
| align="center" style="background:#f0f0f0;"|'''Colony Count <br>(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 | |||
|} | |} |
Latest revision as of 07:40, 13 August 2007
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 |