Difference between revisions of "Team:UMBC-Maryland/Experiments"

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Experiments & Protocols

Agarose Gel

Electrophoresis buffer contents:

10 mL 50x TAE

490 mL deionized water

Contents of Agarose gel:

1g Agarose (1%) or 2g Agarose (2%)

100 mL Electrophoresis buffer

Mix agarose and electrophoresis buffer and microwave for 1.5 minutes.

When the solution is sufficiently cool, add 6 µL of EtBr.

Pour the solution into a bed with a comb levely placed within. Clear the bubbles from the surface and wait for the gel to polymerize.

Mix loading dye and sample in a 1:5 ratio and insert samples into wells. Place 10 µL of DNA ladder into the first well.

Plasmid Extraction

We used NucleoSpin® Plasmid Miniprep kit, Macherey-Nagel according to the manufacturer's instructions for our minipreps.

DNA Assembly and Cloning

We used Gibson Assembly® Protocol (E5510), New England Biolabsaccording to the manufacturer's instructions for cloning.

Restriction Digest

8 μL 2.1 Buffer

5 μL DNA

2 μL EcoR1

2 μL Pst1

Incubate at 37°C for 1 hour

Inoculation Experiments

1. In sterile conditions, prepare 8 flasks with 25 mL LB solution,

What should this page contain?

• Protocols
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Inspiration

• 2014 Colombia
• 2014 Imperial
• 2014 Caltech