Team:EPF-Lausanne/Protocol/DNAConcentration

From 2012.igem.org

(Difference between revisions)
 
(One intermediate revision not shown)
Line 1: Line 1:
<noinclude>{{:Team:EPF-Lausanne/Template/Header}}</noinclude>
<noinclude>{{:Team:EPF-Lausanne/Template/Header}}</noinclude>
-
{{:Team:EPF-Lausanne/Template/ProtocolHeader|DNA concentration measurement|{{{1|}}}}}
+
 
 +
<noinclude>{{:Team:EPF-Lausanne/Template/SetTitle|Nanodrop}}</noinclude>
 +
 
 +
{{:Team:EPF-Lausanne/Template/ProtocolHeader|DNA Concentration Measurement|{{{1|}}}}}
<!--                                                /\            -->
<!--                                                /\            -->
<!--                                                ||            -->
<!--                                                ||            -->
Line 8: Line 11:
<!-- https://2012.igem.org/Team:EPF-Lausanne/Protocol              -->
<!-- https://2012.igem.org/Team:EPF-Lausanne/Protocol              -->
-
* Take a new 6 µl aliquote of the DNA (under the hood) and put back the main DNA tube in the fridge.
+
* Take a 6 µl aliquote of the DNA and put back the main DNA tube in the fridge.
-
* Go to the room by the E. coli lab (not on Friday morning!) with:
+
* Go to the room by the E.Coli lab (LBTM, not on Friday morning!) with:
** The 6 µl aliquote
** The 6 µl aliquote
-
** A 10 µl pipete
+
** A 10 µl pipet
-
** Eventually some 5 µl of TE buffer (they migh have some).
+
** Optionally, the buffer you used for DNA elution (there might be some next to the machine).
* The machine is the NanoDrop Spectrophotometer.
* The machine is the NanoDrop Spectrophotometer.
* On the computer, click on "Nucleic Acid".
* On the computer, click on "Nucleic Acid".
-
* Add 2 µl of (nuclease free) water to the machine's tip as you are asked to and measure.
+
* Put a 2 µl drop of (nuclease-free) water on the machine's tip as you are asked to and measure.
-
* Clean tips (both sides) with a tissue.
+
* Clean tips (both sides) with a quarter of tissue.
-
* Add 2 µl of TE buffer and click on "Blank".
+
* Add 2 µl of the buffer you use and click on "Blank".
-
* Clean tips (both sides) with a tissue.
+
* Clean tips (both sides).
-
* Add 2 µl of DNA and click "Measure".
+
* Add 2 µl of your DNA sample and click "Measure".
* Clean tips (both sides) with a tissue.
* Clean tips (both sides) with a tissue.
-
* Take 2 measurements per sample and average.  
+
* Take 2 measurements per sample (for averaging).
 +
* Print the report when you are done
* Click on exit.
* Click on exit.

Latest revision as of 00:31, 18 September 2012

Protocol: DNA Concentration Measurement


  • Take a 6 µl aliquote of the DNA and put back the main DNA tube in the fridge.
  • Go to the room by the E.Coli lab (LBTM, not on Friday morning!) with:
    • The 6 µl aliquote
    • A 10 µl pipet
    • Optionally, the buffer you used for DNA elution (there might be some next to the machine).
  • The machine is the NanoDrop Spectrophotometer.
  • On the computer, click on "Nucleic Acid".
  • Put a 2 µl drop of (nuclease-free) water on the machine's tip as you are asked to and measure.
  • Clean tips (both sides) with a quarter of tissue.
  • Add 2 µl of the buffer you use and click on "Blank".
  • Clean tips (both sides).
  • Add 2 µl of your DNA sample and click "Measure".
  • Clean tips (both sides) with a tissue.
  • Take 2 measurements per sample (for averaging).
  • Print the report when you are done
  • Click on exit.

The important numbers are:

  • 260/280 ratio, must be > 1.8
  • 260/230 ratio, must be > 2 (too big, > 2.5? , might mean too much salts)
  • Of course the DNA concentration.