Page tree

Versions Compared

Key

  • This line was added.
  • This line was removed.
  • Formatting was changed.

Statistics about an opened sequence can be found on the Statistics tab of the Options Panel. When a region is selected in the sequence, the statistics is interactively re-calculated for this region only. The following information is available:

  • Common statistics (length, molecular weight, etc.) - see the detailed description below
  • Characters occurrence
  • Dinucleotides occurrence - available for nucleotide sequences only

Note that all data, displayed on the Statistics tab, can be selected with the mouse and copied. Use the copy item in the context menu or a shortcut - Ctrl+C on Windows or Linux, Cmd+C on macOS.

...

  • Length
  • GC content
  • Molecular weight
  • Extinction coefficient
  • Melting temperature
  • nmole/OD260
  • μg/OD260

GC content

TODO: add description/meaning?

The percentage of guanine (G) and cytosine (C) bases in the within the sequence or its selected region, for example:

GC-content("ACGTAC") = ((0 + 1 + 1 + 0 + 0 + 1) / 6) * 100% = 50%

If the sequence contains degenerate base characters, average values are used, for example:

GC-content("ACGNBCT") = ((0 + 1 + 1 + 1/2 + 2/3 + 1 + 0) / 7) * 100% ~= 59.52%

In this example "1/2" is used for "N" (any nucleotide), "2/3" us used for "B" (that means "C", "G", or "T" according to the IUPAC notationnucleotide code).

Molecular weight

...

Molecular weight for a single-stranded molecule is calculated as the sum a sum of the atomic masses of the molecule compounds:

DNA molecular weight  = nA*251.24 + nT*242.23 + nC*227.22 + nG*267.24 + (n-1)*

62

61.97

RNA molecular weight = nA*267.

24 

24 + nU*244.20 + nC*243.22 + nG*283.24 + (n-1)*

62

61.97

Here "nA", "nT", "nC", "nG", "nU" denote the number of the corresponding nucleotide in the molecule, "n" is the number of all bases (62 is 61.97 is the weight of an internal phosphate).

Note that for degenerate base characters average value of nucleotide weight is used, for example, if the sequence also contain "Y" characters (that is "C" or "T"), the sum will include one more summand - "nY*(242.23 + 227.22)/2".

Molecular weight for  for a double-stranded molecule is calculated as the sum of the single strands molecular weights.

Extinction coefficient

TODO: add description/meaning?

To calculate the molar extinction  Extinction coefficient, an approach proposed by Richard Owczarzy is used: http://www.owczarzy.net/extinctionDNA.htm. That is for a single-stranded molecule:

Extinction coefficient = sum(extinction coefficients of all dinucleotides) - sum(extinction coefficients of inner mononucleotides)

TODO: should the table below be simplified?

The table below specified the extinction coefficients for dinucleotides and mononulceotides:

DNARNAStack or monomerExtinction coefficientStack or monomer

Extinction coefficientpdA15400pA15400pdC  7400pC  7200pdG11500pG11500pdT  8700pU  9900dApdA27400ApA27400dApdC21200AdC21000dApdG25000ApG25000dApdT22800ApU24000dCpdA21200CpA21000dCpdC14600CpC14200dCpdG18000CpG17800dCpdT15200CpU16200dGpdA25200GpA25200dGpdC17600GpC17400dGpdG21600GpG21600dGpdT20000GpU21200dTpdA23400UpA24600dTpdC16200UpC17200dTpdG19000UpG20000dTpdT16800UpU19600

For example, let's calculate the molar extinction coefficient ("ε") for "ATGCA":

ε(ATGCA) = ε(AT) + ε(TG) + ε(GC) + ε(CA) - ε(T) - ε(G) - ε(C) =

                      = 22800 + 19000 + 17600 + 21200 - 8700 - 11500 - 7400 =

                      = 53000

As for the other statistics, average values are used in case of degenerate base characters.

Extinction coefficientcoefficientfor a double-stranded molecule is calculated as a sum of the extinction coefficients of the two single strandsstrands (es1 + es2) multiplied by coefficient of (1 - hypochromicity h260nm). The hypochromicity effect can be taken into account as follows:

h260nm = (0.287fAT + 0.059fGC)

where fAT and fGC are fractions of AT and GC base pairs, respectively.

Melting temperature

The melting temperature is calculated as follows. For sequences of length 15 or longer:

Tm 64.9 + 41 * (nG + nC - 16.4) / (nA + nT + nG + nC)

For shorter sequences:

Tm = (nA + nT) * 2 + (nG + nC) * 4

Here "nA", "nT", "nC", "nG" denote the number of the corresponding nucleotide.

Context information about a sequence can be found on the All information is contextual, i.e. it shows statistics about the currently selected region (on the selected sequence). The tab includes information about:

...

nmole/OD260

The amount of DNA or RNA represented in nanomoles per 1 unit of absorbance at 260 nm dissolved in 1 ml cuvette with

...

1 cm

...

pathlength.

nmole/OD260 = 1000000 / molarExtCoef

μg/OD260

...

The amount of

...

DNA or RNA represented in microgrames per 1 unit of absorbance at 260 nm dissolved in 1 ml cuvette with

...

1 cm pathlength.

μg/OD260 = nmoleOD260 * molarWeight * 0.001

Amino acid sequence common statistics

...