User:Tara K. Luckau/Notebook/Team ConGen/2011/01/12

Tara's Lab Notebook

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PCR Timbers (by Tara) Extract Sceloporus undulatus continued from 11 January 2011 add 1.5μL RNAse incubate 30 min in 37°C water bath allow to cool to room temperature add 100µL Protein Precipitation Solution; vortex; centrifuge 10 min to pellet transfer supernatant (contains DNA) into new tube add 300µL 100% isopropanol; invert; centrifuge 10 min to pellet pour off supernatant add 300µL 70% ethanol; invert; centrifuge 10 min to pellet pour off supernatant; let air-dry rehydrate with 50-200µL Tris-Cl pH 8.4 (10mM); incubate overnight at room temperature Gel Timbers (by Tara) bands! and they seem to have the same pattern as yesterday's PCR by Rulon (NH 24 has fainter larger band, and smeary smaller band) no big difference between Rulon's or Tara's dNTPs no big difference between Rulon's or Tara's Buffer and MgCl2 conclusion: dNTPs and Buffers are not cause of PCR problems!