User:Beatriz Gimenez De C./Notebook/572/2015/04/01: Difference between revisions

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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</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>}}
|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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==Tasks==
==Tasks==
*
* New protocol
** Vortex each 5 mL test tube of fibers until solution seems homogeneous
** Prepare concentrated buffer -> 50 mM Tris-HCl/20 mM CaCl2 pH 8 buffer.
** Prepare Proteinase K solution
** Add 0.927 mL of the homogeneous fiber sample from the vortexed 5 mL test tube + 0.020 mL of a 2.5 M Tris-HCl/0.5 M CaCl2 pH 8 buffer + 0.053 mL of protease for a total volume of 1 mL into an epitub.
** When the reaction is finished, spin the individual tube for 30 seconds and extract the supernatant for analysis
 
* Run Bradford of Proteinas K for 0, 0.5, 1, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 15, 20, 25, 30, 40, 50 ,60 min.
** 200μM of a 1 in 4 Bradford dilution (1 mL Bradford in 3 mL of Tris/NaCl buffer)
** 150μM of each proteinase K sample
** 550μM of tris/NaCl buffer


==Results==
==Results==

Latest revision as of 00:52, 27 September 2017

Project name Main project page
Previous entry      Next entry

Tasks

  • New protocol
    • Vortex each 5 mL test tube of fibers until solution seems homogeneous
    • Prepare concentrated buffer -> 50 mM Tris-HCl/20 mM CaCl2 pH 8 buffer.
    • Prepare Proteinase K solution
    • Add 0.927 mL of the homogeneous fiber sample from the vortexed 5 mL test tube + 0.020 mL of a 2.5 M Tris-HCl/0.5 M CaCl2 pH 8 buffer + 0.053 mL of protease for a total volume of 1 mL into an epitub.
    • When the reaction is finished, spin the individual tube for 30 seconds and extract the supernatant for analysis
  • Run Bradford of Proteinas K for 0, 0.5, 1, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 15, 20, 25, 30, 40, 50 ,60 min.
    • 200μM of a 1 in 4 Bradford dilution (1 mL Bradford in 3 mL of Tris/NaCl buffer)
    • 150μM of each proteinase K sample
    • 550μM of tris/NaCl buffer

Results



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