User:David Johnston Monje/Notebook/Maize Endophyte Biofertilizers/2013/04/09: Difference between revisions

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==Remains of the Day==
==Remains of the Day==
* Measure spot and speck CFUs by plating (4 half plates will be used) with 50 ul of 10^-3, 10^-4, 10^-5, and 10^-6 --> compare to OD600 values
* Run gel of physical capture TRFLP PCRs and send the lot of them for sequencing after specing the DNA
* Run gel of physical capture TRFLP PCRs and send the lot of them for sequencing after specing the DNA
* Measure spot and speck CFUs by plating (4 half plates will be used) with 50 ul of 10^-3, 10^-4, 10^-5, and 10^-6 --> compare to OD600 values (plated 25 ul of each dilution on a quadrant of TSA and growing overnight at 30 degrees)
** All the dilutions except the  10^-1 were OD600 = 0. At the 10^-1 dilution (10 ul of overnight TSB culture into 90 ul of sterile buffer) Pst T6=0.218, Pst 95=0.319, Pst 08=0.233, Pst 01=0.201, Xg 444=0.274, Xg 1B=0.426, Xg 03=0.022, Xg 00=0.061. Hope I get growth so that I can try to make a curve based on dilutions rather than CFU.  Xg 444 had colonies but was overgrown and difficult to count even at the 10^-8 dilution, while Xg 1B5B had 390 colonies at the 10^-5 dilution and 110 at the 10^-6 dilution. Two days later, Xg 00 yielded 97 colonies for the 10^-5 dilution, 21 for the 10^-6 dilution and 10 for the 10^-7 dilution.
[[image:April 9,2013Gel.tif]]




Revision as of 11:21, 17 April 2013

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Remains of the Day

  • Run gel of physical capture TRFLP PCRs and send the lot of them for sequencing after specing the DNA
  • Measure spot and speck CFUs by plating (4 half plates will be used) with 50 ul of 10^-3, 10^-4, 10^-5, and 10^-6 --> compare to OD600 values (plated 25 ul of each dilution on a quadrant of TSA and growing overnight at 30 degrees)
    • All the dilutions except the 10^-1 were OD600 = 0. At the 10^-1 dilution (10 ul of overnight TSB culture into 90 ul of sterile buffer) Pst T6=0.218, Pst 95=0.319, Pst 08=0.233, Pst 01=0.201, Xg 444=0.274, Xg 1B=0.426, Xg 03=0.022, Xg 00=0.061. Hope I get growth so that I can try to make a curve based on dilutions rather than CFU. Xg 444 had colonies but was overgrown and difficult to count even at the 10^-8 dilution, while Xg 1B5B had 390 colonies at the 10^-5 dilution and 110 at the 10^-6 dilution. Two days later, Xg 00 yielded 97 colonies for the 10^-5 dilution, 21 for the 10^-6 dilution and 10 for the 10^-7 dilution.



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