BIOL 203 Study Guide - Final Guide: Restriction Digest, Restriction Enzyme, Plasmid
Document Summary
Digest a sample of plasmid dna with two restriction enzymes (double digest). Submit: 1 tube of restriction digest and 1 tube of control digest: label two 1. 7ml microcentrifuge tubes: dd double digest & c uncut dna/control (name, Date, section 1: assemble the restriction enzyme digests according to the amounts listed in the table: Incubate only the double digest tube in 37 c dry block for 20 minutes: remove tube from dry block and transfer to the 65 c dry block for 10 minutes. Assemble colony pcr reaction and a control reaction. Gently touch a colony once with the end of the tip. Swirl the colony into the tube of sterile water. Incubate tube in the 100 c heat block for 10 minutes to release dna. Each tube should have a total volume of 20 l: give pcr tubes to ta. Purify plasmid from a 2 ml overnight culture.
