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

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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Applied Soil Microbial Ecology </span>
 
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Applied Soil Microbial Ecology </span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
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|style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}}
 
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* 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^-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.  
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** 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]]
 
[[image:April 9,2013Gel.tif]]
  

Latest revision as of 22:37, 26 September 2017

Owwnotebook icon.png Applied Soil Microbial Ecology Report.pngMain project page
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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.

File:April 9,2013Gel.tif