Team:OU-Norman OK/Project/Notebook

From 2013.igem.org

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<p>TFB2</p>
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<p>TFB2 ph:6.8 (Use KOH to adjust)/ Sterile Filter</p>
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<p>TFB1</p>
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<th>Chemicals</th>
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<th>Mass (g)</th>
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<td>RbCl</td>
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<td>3.02965</td>
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<td>MnCl<sub>2</sub></td>
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<td>1.56996</td>
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<td>Potassium Acetate</td>
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<td> 0.74392</td>
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<td>CaCl<sub>2</sub></td>
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<td>0.27806</td>
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<tr>
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<td>Glycerol</td>
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<td>37.5463</td>
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</table>
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</br>
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<p>TFB2</p>
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<table border = "1" align="center">
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<tr>
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<th>Chemicals</th>
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<th>Mass (g)</th>
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<tr>
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<td>MOPS</td>
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<td>0.53398</td>
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<tr>
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<td>RbCl</td>
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<td>0.30225</td>
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<tr>
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<td>CaCl<sub>2</sub></td>
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<td>2.88320</td>
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<tr>
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<td>Glycerol</td>
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<td>37.5457</td>
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</table>
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Revision as of 23:13, 7 August 2013

Buffers for Preparing Completed E. coli

02/4/13

TFB1: pH: 5.8/ Sterile Filter


Chemicals Concentration (mM)
RbCl 100
MnCl2 50
Potassium Acetate 30
CaCl2 10
Glycerol 15% by Weight

TFB2 ph:6.8 (Use KOH to adjust)/ Sterile Filter

Chemicals Concentrations (mM)
MOPS 10
RbCl 10
CaCl2 75
Glycerol 15% by Weight

TFB1

Chemicals Mass (g)
RbCl 3.02965
MnCl2 1.56996
Potassium Acetate 0.74392
CaCl2 0.27806
Glycerol 37.5463

TFB2

Chemicals Mass (g)
MOPS 0.53398
RbCl 0.30225
CaCl2 2.88320
Glycerol 37.5457
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Making Antibiotic Stocks

02/1/13

Antibiotics are used to isolate organisms into which plasmids containing antibiotic resistance genes have been transformed.

Make antibiotic plates with the following specs.


Antibiotic Concentration (µg/mL) Color Code
Ampicillin 100 Orange
Chloramphenicol 35 Green
Kanamycin 50 Red
Tetracycline 15 Yellow

Stocks should be made at 1000x concentration, so that making 1L of plate medium will require only 1mL of stock antibiotic solution.


Stocks of antibiotics are made at the following concentrations


Ampicillin 100 mg/mL
Chloramphenicol 35 mg/mL
Kanamycin 30 mg/mL
Tetracycline 15 mg/mL

50mL of stock were prepared and split into several 15mL tubes

Stocks should be stored in the refrigerator at 4°C

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01/25/13

Restreaked TOP10 cells on LB plates for isolation of single colonies.

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01/23/13

Sealed TOP10 E. coli cells on LB agar plate. Stored in 37°C incubator

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01/18/13

Poured LB agar plates

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01/16/13

Making Agar Plates

LB agar is used to grow our stocks of Escherichia coli


Recipe: Per 1000 mL

10g Bacto Tryptone 10g

5g Yeast Extract

10g Sodium Chloride (NaCl)

15g Agarose


Mix Components in 1L of dH2O until dissolved. Spilt 1000mL solution into two flask. Cap flask with aluminum to prevent spilling of solution. Autoclave on slow exhaust for 20 minutes. Keep liquid in 65°C water bath to prevent setting until 18-Jan.

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