Difference between revisions of "IGEM:Harvard/2007/Plates"

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(MACS Cell Sort with AIDA Sender Constructs)
 
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Colony Count of Overnight Ligation vs. Shorter (2 hr Ligation)
+
==Colony Count of Overnight Ligation vs. Shorter (2 hr Ligation)==
 
Plated 6-20-07
 
Plated 6-20-07
 
{| {{table}}
 
{| {{table}}
Line 6: Line 6:
 
| align="center" style="background:#f0f0f0;"|'''Colony Count <br>(Short Ligation)'''
 
| align="center" style="background:#f0f0f0;"|'''Colony Count <br>(Short Ligation)'''
 
| align="center" style="background:#f0f0f0;"|'''Colony Count <br>(Overnight Ligation)'''
 
| align="center" style="background:#f0f0f0;"|'''Colony Count <br>(Overnight Ligation)'''
| align="center" style="background:#f0f0f0;"|'''Colony Count <br>(Overweekend Ligation;1uL DNA)'''
+
 
| align="center" style="background:#f0f0f0;"|'''Colony Count <br>(Overweekend Ligation;6uL DNA)'''
 
 
|-
 
|-
 
| OmpA1 + random library, PCR||275 µL||5||41
 
| OmpA1 + random library, PCR||275 µL||5||41
Line 24: Line 23:
 
|-
 
|-
 
| OmpA2 + random library, extension||25 µL||0||1
 
| OmpA2 + random library, extension||25 µL||0||1
 +
|}
 +
 +
== Colony Count of Overweekend Ligation ==
 +
Plated 6/25/07
 +
<i>The following were all done with 275 uL:</i><br>
 +
{|  {{table}}
 +
| align="center" style="background:#f0f0f0;"|'''Cell Type'''
 +
| align="center" style="background:#f0f0f0;"|'''Colony Count <br>(Overweekend Ligation;1uL DNA)'''
 +
| align="center" style="background:#f0f0f0;"|'''Colony Count <br>(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
 +
{|  {{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