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

From OpenWetWare

Jump to: navigation, search
(Autocreate 2013/04/09 Entry for User:David_Johnston_Monje/Notebook/Maize_Endophyte_Biofertilizers)
Current revision (14:21, 17 April 2013) (view source)
(Remains of the Day)
 
(8 intermediate revisions not shown.)
Line 7: Line 7:
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
==Remains of the Day==
==Remains of the Day==
-
* Write up your awesome plans and results here...
+
* 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]]

Current revision

Applied Soil Microbial Ecology Main project page
Previous entry      Next entry

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.

image:April 9,2013Gel.tif



Personal tools