User:Beatriz Gimenez De C./Notebook/572/2015/01/27: 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==
* Produce the lysozyme AuNP fibers.
* Produce the lysozyme AuNP fibers (Made 50, 5 mL samples)
* Weigh the protease
* Weigh 4 mg of Pepsine protease into 8 tubes
* Prepare buffer


==Procedure==
==Procedure==


* Lysozyme-AuNP fibers preparation  
* Lysozyme-AuNP fibers preparation  
** Prepare Lysozyme (0.6 g/L lysozyme) stock solution
** Prepare Lysozyme 20 μM stock solution
*** Measured 0.0261 g of lysozyme into a 100 mL volumetric flask and filled the rest with distilled water
** Prepare 2.5×10<sup>-3</sup> M HAuCl<sub>4</sub> stock solution
** Prepare 2.5×10<sup>-3</sup> M HAuCl<sub>4</sub> stock solution
** Transfer 0.5 mL 2.5×10<sup>-3</sup> M HAuCl<sub>4</sub> into x culture tubes
*** added 0.08539 g of HAuCl4 into a 100 mL volumetric flask and filled the rest with distilled water
*** 0.5×10<sup>-3</sup>(L)*2.5×10<sup>-3</sup>(mol/L)= 1.25 (mol) needed for A 45:1 ratio
** Transfer 0.5 mL 2.5×10<sup>-3</sup> M HAuCl<sub>4</sub> into 50 culture tubes
** Transfer 0.661769570 mL of lysozyme stock solution into x culture tubes
*** 0.5×10<sup>-3</sup>(L)*2.5×10<sup>-3</sup>(mol/L)= 1.25 (mol) needed for a 45:1 ratio
*** 0.6(g/l)/14307(g/mol)= 4.1937510×10<sup>-5</sup>(mol/L)
** Transfer 1.39 mL of lysozyme stock solution into 50 culture tubes
*** 2.77777778×10<sup>-8</sup>(mol)/4.1937510×10<sup>-5</sup>(mol/L)= 0.661769570 mL for a 45:1 ratio
*** 2.77777778×10<sup>-8</sup>(mol)/20*10^-6(mol/ml)= 1.39 mL for a 45:1 ratio
** Fill up culture tubes to 5 mL total with water
** Fill up 50 culture tubes to 5 mL total with distilled water
** Placed in oven at 80ºC for 4h
** Placed in oven at 80ºC for 4h
*Weighed Protease pepsin into 8 tubes, for the next experiment.
** 1. 0.00530 g
** 2. 0.00398 g
** 3. 0.00391 g
** 4. 0.00470 g
** 5. 0.00419 g
** 6. 0.00465 g
** 7. 0.00780 g
** 8. 0.00584 g
* Prepared 5 mM Tris/0.2 M NaCl buffer for reaction of the synthesized lysozyme-AuNP fibers with the proteases
** 1000 mL of buffer was prepared, used 11.68960 g of NaCl + 0.78922 g of Tris in approx. 1000 mL of distilled water.
** 0.5 N HCl added until pH=8.067.




Latest revision as of 00:39, 27 September 2017

Project name Main project page
Next entry

Tasks

  • Produce the lysozyme AuNP fibers (Made 50, 5 mL samples)
  • Weigh 4 mg of Pepsine protease into 8 tubes
  • Prepare buffer

Procedure

  • Lysozyme-AuNP fibers preparation
    • Prepare Lysozyme 20 μM stock solution
      • Measured 0.0261 g of lysozyme into a 100 mL volumetric flask and filled the rest with distilled water
    • Prepare 2.5×10-3 M HAuCl4 stock solution
      • added 0.08539 g of HAuCl4 into a 100 mL volumetric flask and filled the rest with distilled water
    • Transfer 0.5 mL 2.5×10-3 M HAuCl4 into 50 culture tubes
      • 0.5×10-3(L)*2.5×10-3(mol/L)= 1.25 (mol) needed for a 45:1 ratio
    • Transfer 1.39 mL of lysozyme stock solution into 50 culture tubes
      • 2.77777778×10-8(mol)/20*10^-6(mol/ml)= 1.39 mL for a 45:1 ratio
    • Fill up 50 culture tubes to 5 mL total with distilled water
    • Placed in oven at 80ºC for 4h
  • Weighed Protease pepsin into 8 tubes, for the next experiment.
    • 1. 0.00530 g
    • 2. 0.00398 g
    • 3. 0.00391 g
    • 4. 0.00470 g
    • 5. 0.00419 g
    • 6. 0.00465 g
    • 7. 0.00780 g
    • 8. 0.00584 g
  • Prepared 5 mM Tris/0.2 M NaCl buffer for reaction of the synthesized lysozyme-AuNP fibers with the proteases
    • 1000 mL of buffer was prepared, used 11.68960 g of NaCl + 0.78922 g of Tris in approx. 1000 mL of distilled water.
    • 0.5 N HCl added until pH=8.067.



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