DEPARTMENT OF PATHOLOGY
Reverse Complement and
Sequence Tm Utility Alexa Barnes
June 2012
DEPARTMENT OF PATHOLOGY
Introduction
The Reverse Complement and Sequence Tm Utility
•  Calculates the reverse, complement, and reverse
complement of an entered sequence
• Calculates the Tm
• Provides a double stranded visualization in real time.
• Allows for the addition of mismatches and updates the Tm.
DEPARTMENT OF PATHOLOGY
Basic Use
Type or copy and paste
sequence here:
Reverse, complement,
and reverse complement
automatically populate as
entered in real time.
DEPARTMENT OF PATHOLOGY
Tm Calculation
Click on for more
information
Conditions can
be changed
DEPARTMENT OF PATHOLOGY
How to Enter a Mismatch
1
Highlight a single
nucleotide using the
mouse
2
Replace with lower
cased mismatched
nucleotide
3
Double stranded view
updates to show
mismatch with an “x”
DEPARTMENT OF PATHOLOGY
References
[ free Magnesium ] according to :
von Ahsen N, Wittwer CT, Schütz E. “Oligonucleotide melting temperatures under PCR
conditions: nearest-neighbor corrections for Mg(2+), deoxynucleotidetriphosphate, and dimethyl
sulfoxide concentrations with comparison to alternative empirical formulas”. Clin Chem. 2001
Nov;47(11):1956-61.
[ Monovalent ], Mismatches and tetrad thermodynamics according to :
SantaLucia J Jr, Hicks D. “The thermodynamics of DNA structural motifs”. Annu Rev Biophys
Biomol Struct. 2004;33:415-40.