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

(Difference between revisions)
 Revision as of 17:26, 9 April 2013 (view source) (→Remains of the Day)← Previous diff Revision as of 10:54, 10 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 444=0.426, Xg 444=0.022, Xg 444=0.061. Hope I get growth so that I can try to make a curve based on dilutions rather than OD. + ** 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 OD. [[image:April 9,2013Gel.tif]] [[image:April 9,2013Gel.tif]]

## Revision as of 10:54, 10 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 OD.