Left, Tetracycline, and Right Overlap Update
These past couple weeks in lab have been very interesting. First, we decided to give the right fragment of deinococcus radiodurans and tetracycline from E. coli a try. We are doing this because we haven’t seen much success in adding our right fragment to our left and tet fragments. We initially had isolated more right fragment via gel excision. We had performed two different right isolations. Our ng/μL reading on R 1 isolation was 18.8 ng/μL, and for our R 2 isolation it was 27.7ng/μL. The sample we chose for our overlap reaction was R 2 . We then decided to do an 8-sample overlap heat gradient of right fragment and tetracycline from 35 °C to 60°C. The recipe we used is below: 11μL 7μL 2μL 4μL Master Mix PCR H 2 0 Right Fragment Tetracycline Fragment From L to R: 60°C, 57.9°C, 54.6°C, 50.1°C, 44.5°C, 39.6°C, 36.7°C, 35°C The two best bands we saw were at 57°C and 35°C. We gel excised these bands by loading the rest of our sample onto a gel. With these