Team:UCLA/Notebook/Spider Silk Genetics/1 May 2015

iGEM UCLA





5/1/2015

5/1/2015


1 Calculation for Primer exhaustion

∙ Vector+insert = 2205 bp (150 bp insert) (2055 bp vector)

∙ DNA: 649 g/mole/bp

∙ AB: 143105 g/mole

∙ 500 pg / 1431045 g/mol = 3.494 E -16 moles DNA

∙ 1.25 uL of 10 uM primers = 1.25 E -11 moles primers

∙ 1.25 E -11 / 3.494 E -16 = 35775.6

∙ log2(35775.6) = 15.1 cycles (assuming perfect PCR)

2 Modified PCR using post-elution primers

Redo PCR using same amounts of reagents, but with 20 cycles instead of 30. Made 2x 50 uL for each MaSp plasmid.