Prbbbb:dna measurement on plate v1

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(Calculation)
(Calculation)
Line 21: Line 21:
* path length:
* path length:
   d = V / (3.3^2 * mm^2)  
   d = V / (3.3^2 * mm^2)  
-
  V... volume per well in µl
+
V... volume per well in µl
'''formula'''
'''formula'''
-
c = (A * e) / d
+
  c = (A * e) / d
* c... DNA concentration in ng / µl
* c... DNA concentration in ng / µl

Revision as of 07:02, 20 May 2009

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Contents

Overview

Materials

  • Tecan plate reader
    • (or any reader that accepts 384 plates and can measure at 260 and 280 nm)
  • 384 well UV transmissive plate with flat clear bottom
    • we use Greiner UV star Cat # 781 801
  • miniprepped DNA, at least 30 µl

Calculation

parameters

  • well dimension: flat bottom 3.3 mm x 3.3 mm
  • DNA extinction coefficient e: 50 [(ng * cm) / µl]
  • path length:
 d = V / (3.3^2 * mm^2) 

V... volume per well in µl

formula

 c = (A * e) / d
  • c... DNA concentration in ng / µl
  • A... Absorbance @ 260 nm
  • d... path length in cm

Procedure

  1. Pipette exact volume of DNA samples into 384 well plate
  2. measure 260 nm and 280 nm absorbance on plate reader
  3. Pipette sample back into storage plate
  4. calculate DNA concentration from formula above (compare to reference)


Notes

Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!

raik: This protocol is quick and robust in my hands.


References

Contact

or instead, discuss this protocol.

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