User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/01/29: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
(Autocreate 2013/01/29 Entry for User:Madeleine_Y._Bee/Notebook/Single_Molecule_Fluorescence)
 
(fix raw html notebook nav)
 
(2 intermediate revisions by one other user not shown)
Line 2: Line 2:
|-
|-
|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}}}}]]}}
|-
|-
| colspan="2"|
| colspan="2"|
<!-- ##### 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. ##### -->
==Entry title==
==January 29, 2013==
* Insert content here...
*Concentration calculations-
1) make 100μL of 1μM 4 strand DNA:<br>
2.656μL 4 strand DNA in 97.344μL filtered H<sub>2</sub>O <br>
2) make 100μL of 1μM 8 strand DNA:<br>
1.428μL 8 strand DNA in 98.527μL filtered H<sub>2</sub>O <br>
3) make 6.49nM Molecular Beacon: <br>
731nM(V)=6.49nM(1000μL) <br>
V=8.878μL MB in 991.12175μL unfiltered H<sub>2</sub>O <br>
4) make TRIS buffer (pH of 7.2): <br>
(121.14g/mol TRIS)(5mmol/L)(1mol/1000mmol)(50mL)(1L/1000mL)=0.030285g TRIS  in 50mL solution <br>
*combine 0.0352g TRIS, 9 drops HCl, 5mL distilled H<sub>2</sub>O, 0.0025g TRIS, 20mL H<sub>2</sub>O <br>
*result: 5mM TRIS buffer solution with a pH of 7.2 <br>
5) make 3.5μM ThT <br>
(318.86g/mol ThT)(3.5mmol/L)(1mol/10^3mmol)=1.116g/L(1L/1000mL)(10mL)=0.01116g ThT in 10mL solution <br>
*dissolve 0.0134g ThT (massed value, approximated for 0.01116g) in 10mL filtered H<sub>2</sub>O : 3.5mM ThT solution <br>
*dilute 10μL 3.5mM ThT solution in 9990μL filtered H<sub>2</sub>O : 3.5μM ThT solution <br>
6) make DNA solutions at varying concentrations: <br>
DNA (4 strand) concentration: 265.6μM <br>
DNA (8 strand) concentration: 142.8μM <br>
*1:265.6(V)=3.5(250μL) <br>
V=3.29μL 4 strand DNA in 246.71μL TRIS buffer solution <br>
*2:2.82μL 4 strand DNA in 247.18μL TRIS buffer solution <br>
*3:2.35μL 4 strand DNA in 247.65μL TRIS buffer solution <br>
*4:1.88μL 4 strand DNA in 248.12μL TRIS buffer solution <br>
*5:1.41μL 4 strand DNA in 248.59μL TRIS buffer solution <br>
*6:142.8(V)=3.5(250μL)<br>
V=6.13μL 8 strand DNA in 243.87μL TRIS buffer solution <br>
*7:5.25μL 8 strand DNA in 244.75μL TRIS buffer solution <br>
*8:4.38μL 8 strand DNA in 245.62μL TRIS buffer solution <br>
*9:3.50μL 8 strand DNA in 246.50μL TRIS buffer solution <br>
*10:2.63μL 8 strand DNA in 247.38μL TRIS buffer solution <br>
7)mix 100μL of each of the 10 DNA solutions with 100μL 3.5μM ThT
 




Latest revision as of 22:25, 26 September 2017

Project name Main project page
Next entry

January 29, 2013

  • Concentration calculations-

1) make 100μL of 1μM 4 strand DNA:
2.656μL 4 strand DNA in 97.344μL filtered H2O
2) make 100μL of 1μM 8 strand DNA:
1.428μL 8 strand DNA in 98.527μL filtered H2O
3) make 6.49nM Molecular Beacon:
731nM(V)=6.49nM(1000μL)
V=8.878μL MB in 991.12175μL unfiltered H2O
4) make TRIS buffer (pH of 7.2):
(121.14g/mol TRIS)(5mmol/L)(1mol/1000mmol)(50mL)(1L/1000mL)=0.030285g TRIS in 50mL solution

  • combine 0.0352g TRIS, 9 drops HCl, 5mL distilled H2O, 0.0025g TRIS, 20mL H2O
  • result: 5mM TRIS buffer solution with a pH of 7.2

5) make 3.5μM ThT
(318.86g/mol ThT)(3.5mmol/L)(1mol/10^3mmol)=1.116g/L(1L/1000mL)(10mL)=0.01116g ThT in 10mL solution

  • dissolve 0.0134g ThT (massed value, approximated for 0.01116g) in 10mL filtered H2O : 3.5mM ThT solution
  • dilute 10μL 3.5mM ThT solution in 9990μL filtered H2O : 3.5μM ThT solution

6) make DNA solutions at varying concentrations:
DNA (4 strand) concentration: 265.6μM
DNA (8 strand) concentration: 142.8μM

  • 1:265.6(V)=3.5(250μL)

V=3.29μL 4 strand DNA in 246.71μL TRIS buffer solution

  • 2:2.82μL 4 strand DNA in 247.18μL TRIS buffer solution
  • 3:2.35μL 4 strand DNA in 247.65μL TRIS buffer solution
  • 4:1.88μL 4 strand DNA in 248.12μL TRIS buffer solution
  • 5:1.41μL 4 strand DNA in 248.59μL TRIS buffer solution
  • 6:142.8(V)=3.5(250μL)

V=6.13μL 8 strand DNA in 243.87μL TRIS buffer solution

  • 7:5.25μL 8 strand DNA in 244.75μL TRIS buffer solution
  • 8:4.38μL 8 strand DNA in 245.62μL TRIS buffer solution
  • 9:3.50μL 8 strand DNA in 246.50μL TRIS buffer solution
  • 10:2.63μL 8 strand DNA in 247.38μL TRIS buffer solution

7)mix 100μL of each of the 10 DNA solutions with 100μL 3.5μM ThT



Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:Madeleine_Y._Bee/Notebook/Single_Molecule_Fluorescence/2013/01/29&oldid=1011929"