BIOLOGY 173 Lecture Notes - Lecture 10: Electrophoresis, Lysozyme, P200
Document Summary
Molecular biology lab: clean pcr product, cut pcr product and cloning vector with the same restriction enzyme, heat-inactivate the restriction enzyme, mix and ligate pcr fragment with vector. Take gfp gene from pcr product and insert into cloning vector. Run leftover pcr product on gel to size. Pcr cleanup (to get rid of excess primer/ntps which might interfere with digestion or ligation) Longer dna fragments (100+ bp) will remain bound to the column matrix while primers, nucleotides, etc. will flow through: discard the liquid in the collection tube, but keep the tube. Put the column back in the collection tube: wash the column by adding 750 l pe buffer. Centrifuge for 1 minute at maximum speed: discard the flow-through and put the column back in the collection tube, centrifuge for 1 minute at maximum speed. This will ensure that the column is dry: put the column into a clean 1. 5 ml microcentrifuge tube.
