User:David Johnston Monje/Notebook/Maize Endophyte Biofertilizers/2013/04/09: Difference between revisions
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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"| | |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]] }}{{#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. | ** 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
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