IGEM:Harvard/2007/Plates

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Current revision (10:40, 13 August 2007) (view source)
(MACS Cell Sort with AIDA Sender Constructs)
 
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== Colony Count of Electroporation ==
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Plated 7/11/07
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{|  {{table}}
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| align="center" style="background:#f0f0f0;"|'''Plate'''
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| align="center" style="background:#f0f0f0;"|'''Dilution of cells'''
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| align="center" style="background:#f0f0f0;"|'''0ng DNA'''
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| align="center" style="background:#f0f0f0;"|'''25ng DNA'''
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| align="center" style="background:#ff0000;"|'''50ng DNA'''
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| align="center" style="background:#f0f0f0;"|'''100ng DNA'''
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| align="center" style="background:#f0f0f0;"|'''250ng DNA'''
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| align="center" style="background:#f0f0f0;"|'''500ng DNA'''
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|-
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| LB-KAN||10^-2|| || ||521||136||197||361
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|-
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| LB||10^-3||489||329||576||267||130||338
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|}
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Cell Type'''
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| align="center" style="background:#f0f0f0;"|'''Volume Plated'''
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| align="center" style="background:#f0f0f0;"|'''Colony Count <br>(white)'''
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| align="center" style="background:#f0f0f0;"|'''Colony Count <br>(red)'''
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|-
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| AIDA1 + strep, MACS||5 µL||5||0
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|-
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| AIDA1 + strep, MACS||50 µL||31||5
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|-
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| AIDA1 + strep, MACS||300 µL||235||10
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|-
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| AIDA1 + his, MACS||5 µL||0||1
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|-
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| AIDA1 + his, MACS||50 µL||18||0
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|-
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| AIDA1 + his, MACS||300 µL||206||11
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|-
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==MACS and FACS Cell Sort with AIDA Construct==
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Plated 7-20-07
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''Cell Type'''
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| 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)'''
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|-
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| AIDA1 + strep, (+) fraction, FACS||10 µL||0||19
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|-
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| AIDA1 + strep, (-) fraction, FACS||10 µL||0||18
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|-
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| AIDA1 + his, (+) fraction, FACS||10 µL||187||16
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|-
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| AIDA1 + his, (-) fraction, FACS||10 µL||0||12
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|}
 +
 +
==MACS Cell Sort with AIDA Construct-All Induced==
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Plated 8-5-07
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{| {{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)'''
 +
 +
|-
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| AIDA1 + strep, (+) fraction, MACS||120 µL||101||3
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|-
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| AIDA1 + strep, (+) fraction, MACS||25 µL||13||2
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|-
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| AIDA1 + his, (+) fraction, MACS||120 µL||29||22
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|-
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| AIDA1 + his, (+) fraction, MACS||25 µL||5||4
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|}
 +
 +
==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)'''
 +
 +
|-
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| AIDA1 + strep sender, (+) fraction, MACS||75 µL||325||24
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|-
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| AIDA1 + strep sender, (+) fraction, MACS||20 µL||103||8
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|-
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| AIDA1 + his sender, (+) fraction, MACS||75 µL||588||49
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|-
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| AIDA1 + his sender, (+) fraction, MACS||20 µL||317||30
|}
|}

Current revision

Contents

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, PCR275 µL541
OmpA1 + random library, PCR25 µL02
OmpA1 + random library, extension275 µL030
OmpA1 + random library, extension25 µL00
OmpA2 + random library, PCR275 µL01
OmpA2 + random library, PCR25 µL00
OmpA2 + random library, extension275 µL1277
OmpA2 + random library, extension25 µL01

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/S0127
OmpA1/Group1/S2824
OmpA1/Group2/B10119
OmpA1/Group2/S2479

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-KAN10^-2 521136197361
LB10^-3489329576267130338


MACS and FACS Cell Sort with AIDA Construct

Plated 7-20-07

Cell Type Volume Plated Colony Count
(white)
Colony Count
(red)
AIDA1 + strep, MACS5 µL50
AIDA1 + strep, MACS50 µL315
AIDA1 + strep, MACS300 µL23510
AIDA1 + his, MACS5 µL01
AIDA1 + his, MACS50 µL180
AIDA1 + his, MACS300 µL20611
Cell Type Volume Plated Colony Count
(white)
Colony Count
(red)
AIDA1 + strep, (+) fraction, FACS10 µL019
AIDA1 + strep, (-) fraction, FACS10 µL018
AIDA1 + his, (+) fraction, FACS10 µL18716
AIDA1 + his, (-) fraction, FACS10 µL012

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, MACS120 µL1013
AIDA1 + strep, (+) fraction, MACS25 µL132
AIDA1 + his, (+) fraction, MACS120 µL2922
AIDA1 + his, (+) fraction, MACS25 µL54

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, MACS75 µL32524
AIDA1 + strep sender, (+) fraction, MACS20 µL1038
AIDA1 + his sender, (+) fraction, MACS75 µL58849
AIDA1 + his sender, (+) fraction, MACS20 µL31730
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