Difference between revisions of "Team:RPI Troy NY/InterLab"

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<center>Chemical Transformation<center>
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<th><h5>Introduction</h5></th>
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<td><p>Use RuCl2 from -80 freezer</p></td>
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<th><h5>Materials</h5></th>
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<td><p> 100uL cells (thawed on ice)
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<p>  250uL sterile SOC
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<th><h5>Procedure</h5></th>
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<td><p>Process will take approximately 1h45min
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<p>  Add DNA to thawed cells on ice
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<p>  30 minute incubation on ice for transformation
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<p>  42oC heat shock for 45 seconds
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<p>  1 minute on ice
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<p>   Add 250mL sterile SOC to 2mL tube
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<p>  Place in 37oC incubator at 250 RPM for 1 hour (recovery)
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Revision as of 00:32, 31 October 2017

Home Team Collaborations Project Results
Interlab Study Safety Human Practices Attributions

Interlab Study

We are very excited to announce we've transformed our DNA parts for the 2017 iGEM Interlab Study. Check out our Twitter page for weekly updates on our project!

We will be using DNA parts from Plate 7 in the distribution kit and are looking forward to collecting our first round of data.

♦Process Flow♦

 Steps with an asterisk are not required in the iGEM interlab study, rather they are control steps to preserve the DNA and ensure that if anything goes wrong, we have a stock to retransform

 We choose two colonies per plate so we can run biological duplicates. This can provide information on variability of compounds within a single organism (in this case, DH5a e. Coli)

 The iGEM plate reader protocol has a series of dilutions, so we will have to pay careful attention to those details when the time comes

 Form 1 can be filled out at any time, Form 2 requires spreadsheet data from plate reader, Form 3 is in regard to the culturing process

♦Protocols♦

LB Agar Plates
Introduction

Used in conjunction with appropriate antibiotics to selectively plate
Materials

 LB Agar (NOT agarose), 35g/L

 appropriate antibiotic (ie, for 800mL LB Agar, add 800uL Amp, or 400Cm)

Procedure

Stir bar before autoclave

 If stir bar is forgotten, sterilize in EtOH to flame before dropping in autoclaved LB agar

 Mix appropriate amount of agar and DI water, dissolving completely to prevent burning agar

 Autoclave on liquid cycle (15 min is fine)

 Immediately after completing autoclave, cool on stir plate until not too hot to hold

 Add antibiotic

 Don't degrade antibiotic, but don't let cool too much and harden

Chemical Transformation
Introduction

Use RuCl2 from -80 freezer
Materials

 100uL cells (thawed on ice)

 250uL sterile SOC

Procedure

Process will take approximately 1h45min

 Add DNA to thawed cells on ice

 30 minute incubation on ice for transformation

 42oC heat shock for 45 seconds

 1 minute on ice

 Add 250mL sterile SOC to 2mL tube

 Place in 37oC incubator at 250 RPM for 1 hour (recovery)