Title: Restriction Digest Verification of Recombinant bglA-pLATE31 Plasmid-2
Course: BTEC-1100-301-302-303-F21 > Section: Module 4 - Applied Molecular Biology

Below is the table you will fill out and use for the digest of your recombinant plasmid.

You will need to obtain the concentration of your purified plasmid using the spectrophotometer and digest 100 ng of your product.

Incubate the reactions for 30 minutes at 37°C.

You should be familiar already with the steps for running a gel to see if your digests were successful.

Make a 0.8% agarose gel by adding 0.4 grams of agarose to 50 ml of 1X TAE. After dissolving the gel add 5 μl of 10 mg/ml ethidium bromide for visualization.

You will need to cast a gel with a 6-well comb.

You will add 6 µl of 6X loading dye to your reactions and run the entire 36 µl on the gel.

Don’t forget to run 5 μl of the DNA ladder.