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

From OpenWetWare
Jump to navigationJump to search
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 but were overgrown and difficult to count even at the 10^-8 dilution.  
** 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^-7 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 11:12, 17 April 2013

Applied Soil Microbial Ecology <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

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^-7 dilution. Two days later, Xg 444 and Xg 1B5B had colonies but were overgrown and difficult to count even at the 10^-8 dilution.



Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:David_Johnston_Monje/Notebook/Maize_Endophyte_Biofertilizers/2013/04/09&oldid=692361"