Transformation of last day: four plates without ampicillin have growth; four plates with ampicillin have growth except in the control expected.

Nanodrop of pPink-HC:

*Measurement 1: 444 ng/ul, 1.56 (260/280), 2.03 (260/230)
*Measurement 2: 393.4 ng/ul, 1.60 (260/280), 1.91 (260/230)

Plate count (results):

puc19 + amp with a little but growth (so incubate one more day)
GrFP + Amp= 2468 ufc and pPink +Amp= 2138 ufc
Other plates were non-accounting
Efficiency of puc19 transformation (2 days)= (70 ufc)(551ul)/(100ul)= 3.857X106

Enzymatic Digestion: Used pPink-HC, pPink-HC + XbaI+ sacI, puc19; and used NEB cutter of new England Biolabs. Buffers of promega for these enzymes (we used buffer E)

Reaction Digestion (20ul): NF water (12.8 ul), buffer (1X) (2ul), BSA (0.1X) (0.2ul); Plasmid pPink-HC (3ul); Enzyme sacI (1ul); Enyme XbaI (1ul).

Gel Electrophoresis: Prepared the gels such that it is 0.7% agarose. Weigh out approximately for 0.525g of agarose (see protocol for Electrophoresis gel).

Competent cells preparation: Inoculate 10ul TOP10F stock + 20ml de LB; and prepare LB medium liquid (100mL) and solid (200mL).

Team:Tec-Monterrey/allergen/notebook

From 2012.igem.org