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

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 * style="background-color: #BCED91"|[[Image:owwnotebook_icon.png|128px]] Tara's Lab Notebook
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PCR Timbers (by Tara)

 * [[Image:PCROpt_Timbers.jpg|800 px]]

Extract Sceloporus undulatus

 * continued from 11 January 2011
 * 1) add 1.5μL RNAse
 * 2) incubate 30 min in 37°C water bath
 * 3) allow to cool to room temperature
 * 4) add 100µL Protein Precipitation Solution; vortex; centrifuge 10 min to pellet
 * 5) transfer supernatant (contains DNA) into new tube
 * 6) add 300µL 100% isopropanol; invert; centrifuge 10 min to pellet
 * 7) pour off supernatant
 * 8) add 300µL 70% ethanol; invert; centrifuge 10 min to pellet
 * 9) pour off supernatant; let air-dry
 * 10) rehydrate with 50-200µL Tris-Cl pH 8.4 (10mM); incubate overnight at room temperature

Gel Timbers (by Tara)

 * [[Image:20110112_TimberTKL.tif|800 px]]
 * 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!


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