User:David Johnston Monje/Notebook/Maize Endophyte Biofertilizers/2011/11/17

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Remains of the Day

  • Going to extract RNA from the rest of the leaf samples from the potato induction experiment, make cDNA, then later today do a PCR on all samples (30 cycles, 56 degree anealing) to see if IBR had any effect.
  • Gonna repeat induction protocol on tomatoes - so treat tommorow, wait 3 days, repeat, then harvest on day 6.
  • As ITS primers appear to work exponentially better on pure cultures, than on mixed, Im gonna have to explore how to improve signal from soil.
  • Good review paper about mycorhizal fungi TRFLP: http://www.springerlink.com/content/r15h4522615r25m4/fulltext.html

Used less stringent conditions and got more product, even on TVD samples. Unfortunately, not sure that this represents soil - Amy thinks it represents plant root DNA, and as such may cause spurious amplification of plant 18S rather than fungus. Hmm...

300pix


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