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

(Difference between revisions)
 Revision as of 10:00, 16 April 2013 (view source) (→Remains of the Day)← Previous diff Revision as of 10:36, 16 April 2013 (view source) (→Remains of the Day)Next diff → Line 9: Line 9: * 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) * 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. For Xg 00, it yielded 97 colonies for the 10^-5 dilution, 21 for the 10^-6 dilution and 10 for the 10^-6 dilution. Two days later, Xg 444 and Xg 1B5B had colonies - (Xg 444 at 10^-7=52, 10^-8=7) and (Xg 1B5B at 10^-7=11, 10^-8=5). + ** 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. For Xg 00, it yielded 97 colonies for the 10^-5 dilution, 21 for the 10^-6 dilution and 10 for the 10^-6 dilution. Two days later, Xg 444 and Xg 1B5B had colonies but were overgrown and difficult to count even at the 10^-8 dilution. [[image:April 9,2013Gel.tif]] [[image:April 9,2013Gel.tif]]

Revision as of 10:36, 16 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. For Xg 00, it yielded 97 colonies for the 10^-5 dilution, 21 for the 10^-6 dilution and 10 for the 10^-6 dilution. Two days later, Xg 444 and Xg 1B5B had colonies but were overgrown and difficult to count even at the 10^-8 dilution.