Week+of+December+12

__December 12th__ Today, we did new reactions with the PCR reagents that were proveded by Genotyp. On Wednesday, we will run a gel, and then extract the vector and GFP so that we can nanodrop them. Hopefully, we get good results.

__December 14th__ Today we ran the tests that we had planned on Monday. However, the nanodrop showed that we only had contamination. This has now happened multiple times. Therefore, we think something may be wrong with the extraction kit.

Procedure for DNA Extraction from Agraose gel: (From Qiagen) (Image from Google Images)
 * Excise DNA fragment from agarose gel with clean, sharp scalpel.
 * Weigh gel slice in colorless tube. (Ours came to about 300mg each.) Add 3 volumes of Buffer QG to 1 volume of gel (100mg ~ 100µl)
 * Incubate @ 50C for 10 min.
 * SOLUBILIZE AGAROSE COMPLETELY
 * To help dissolve gel, vortex tube every 2-3 min during incubation.
 * After the gel has dissolved, check that the color of the mixture is yellow. If not, add 10µl 3M NaOAc. Or just add the NaOAc anyway.
 * Add 1 gel volume of isopropanol to the sample and mix
 * Do only if 4kb
 * Place spin column in 2mL collection tube.
 * To bind DNA, apply sample to column. Centrifuge 1 min.
 * Discard flowthrough and place column back in same collection tube.
 * Recommended: Add 0.5mL of Buffer QG to column and centrifuge for 1 min. ( The column would not allow for all of the wash to be added at once; we did multiple washes.)
 * To wash, add 0.75mL of Buffer PE to column and centrifuge for 1 min.
 * Discard flowthrough and centrifuge for an additional 1 min @ 17900 x g (13000rpm).
 * Place column into a clean 1.5mL microcentrifuge tube.
 * To elute DNA, add 30µL Buffer EB to center of membrane, let column stand for 1 min, then centrifuge for 1 min