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.
Restriction Digest
8 μL 2.1 Buffer
5 μL DNA
2 μL EcoR1
2 μL Pst1
Incubate at 37oc for 1 hour
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