Team:UNICAMP-EMSE Brazil/Notebook/23 September 2011

Electrophoresis Gel 1, 23/09/2011.
Gel 1: (1-3) Const_prom+RBS+QseB+RBS+QseC+Term+flhDC_prom+RBS+GFP+HlyA +T / (4) empty / (5-8) SoxS+RBS+IL-10+HlyA+T / (9) empty / (10-15) Const_prom+RBS+SoxR+T +SoxS_prom+RBS+GFP+HlyA+T / (16) empty / (17-18) Const_prom+RBS+QseB+RBS+QseC+Term (pSB1C3 vector)

23 September 2011

Finishing constructs and shipping sequences

Quantification and checking digestion of sequences

Objectives:

1. Digestion with E-P to check previously transformed constructions

Samples:

• Gel 1:

• (1-3) Const_prom+RBS+QseB+RBS+QseC+Term+flhDC_prom+RBS+GFP+HlyA +T / (4) empty / (5-8) SoxS+RBS+IL-10+HlyA+T / (9) empty / (10-15) Const_prom+RBS+SoxR+T +SoxS_prom+RBS+GFP+HlyA+T / (16) empty / (17-18) Const_prom+RBS+QseB+RBS+QseC+Term (pSB1C3 vector)

• Ladder:

• 1kB Plus DNA Ladder (Invitrogen)

• Gel Agarose concentration:

• 1,0%

• Result:

• (1-3) Ok. All three samples have inserts with the expected size (~3400bp). Vector: Const_prom (iGEM)
• (5-8) None of the vectors have an insert with the expected (~970bp). Vector: Terminator (iGEM)
• (10-15) All inserts have similar sizes, a little bit smaller than expected (~1800bp). Vector: Const_prom (iGEM)
• (17-18) Only the right sample (C4) have the expected size (~2100bp), which is near to the expected size of the vector (pSB1C3), barely separated.