User:Anthony Salvagno/Notebook/Research/2009/10/30/Primer Melting Conditions

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I want to figure out the Tm for my primers.

Tools

Manipulate a DNA Sequence
mFold via Integrated DNA Technologies This is pretty easy to use.
mFold via RPI
DINAMelt This is what I am using for analysis here.

Approach

I have 4 oligos to compute the melting Temp (Tm) of. Into mFold I am inputting these parameters:

  • DNA sequence - there are two boxes. The left input is my oligo and the right input is the inverse compliment.
  • 50mM Na+
  • 2.5mM MG++
  • 10uM oligo concentration
  • Temp 37C

Results

SJK 18:09, 30 October 2009 (EDT)

18:09, 30 October 2009 (EDT)
Sweet, they all look within similar range.

F853D

Tm = 68.2C

F834D

Tm = 67.2C

R2008

Tm = 66.2C

R1985

Tm = 69.6C

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