User:David Johnston Monje/Notebook/Maize Endophyte Biofertilizers/2012/01/16

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Remains of the Day

  • Do 50 ul PCR on DNA. Use 1 ul of undilute DNA, modified Amy recipe, but with labelled 799F and 1492R primers and IT1F and ITS4. Use 50 degree annealing temperature and 30 cycles.

dH2O - 37.5 ul

10X Buffer - 5 ul

MgCl2 (50 mM) - 2 ul

BSA (20 mg/mL) - 1.25 ul

dNTPs (10 mM) - 1 ul

Primer F (10 mM) - 1 ul

Primer R (10 mM) - 1 ul

Taq Polymerase - 0.25 ul

Total - 50 ul

PCR program is 2 mins 95, 30 cycles (30 sec 95, 30 sec @50 degrees, 1 mins 72 degrees), 7 mins @ 72 dgrees

  • Spray potatoes again



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