R : How To Compute The Melting Temperature Tm
4
2
Entering edit mode
12.0 years ago

Hi,

Is there a R package to compute the Tm of a dna sequence ?

Thanks,

N.

r • 8.8k views
ADD COMMENT
4
Entering edit mode
4.0 years ago
manojmonmax ▴ 90

Hello,

There is a simple formula to calculate the melting temperature of a primer
  • If you have a sequence that is less than 14 nucleotides, then you can use the formula :

Melting temperature(°C) = (nA+nT) 2 + (nG+nC) 4

  • If you have a sequence that is greater than 14 nucleotides, then you can use the formula :

Melting temperature(°C) = 64.9 + 41 * (nG+nC-16.4)/(nA+nT+nG+nC)

where,

  • nA : Number of adenine bases in the sequence
  • nT : Number of thymine bases in the sequence
  • nC : Number of cytosine bases in the sequence
  • nG : Number of guanine bases in the sequence
ADD COMMENT
2
Entering edit mode
12.0 years ago
JC 13k

I'm not aware of a package with the formula, but you can use the the Wallace rule for an approximation in short sequences (primers):

Tm = 2°C(A+T) + 4°C(G+C)

Or the Howley formula for longer sequences:

Tm = 81.5 + 0.4m1(%GC) – 500/L + 16.6 log[M]

where L refers to the length of the oligonucleotide, and [M] is the concentration of monovalent cations.

ADD COMMENT
1
Entering edit mode
12.0 years ago
KCC ★ 4.1k

If you know python (or can manage to read it reasonably well), then you can translate this function into R: http://biopython.org/DIST/docs/api/Bio.SeqUtils.MeltingTemp-pysrc.html#Tm_staluc

It's not a very lengthy function, and would probably not take very long.

ADD COMMENT
0
Entering edit mode

Hi, How to get the latest Biopython with the current version of Tm_staluc? As my matchine is a old version,it cannot use of function of Tm_NN. Thanks

ADD REPLY
0
Entering edit mode
12.0 years ago

Once upon a time I wrote a simple Perl script that will do this. Similar to the Python script suggested by 'George' this script uses a nearest neighbor approach. The nearest neighbor and thermodynamic calculations are done essentially as described by Breslauer et al., (1986) Proc. Nat. Acad. Sci. 83:3746-50 but using the thermodynamic data values published by Sugimoto et al., (1996) Nucl. Acids Res. 24:4501-4505. It assumes that the sequences are not symmetric and contain at least one G or C. The minimum length for the query sequence is 8.

Converting it to R should not be too painful hopefully.

getNearestNeighborTm.pl

ADD COMMENT

Login before adding your answer.

Traffic: 2306 users visited in the last hour
Help About
FAQ
Access RSS
API
Stats

Use of this site constitutes acceptance of our User Agreement and Privacy Policy.

Powered by the version 2.3.6