Method of characterizing polynucleotides
First Claim
1. A method of characterizing polydispersed, single-stranded or double-stranded polynucleotides according to their molecular size distribution comprising the steps ofconverting the polynucleotides to single-stranded polynucleotides,electrophoresing the single-stranded polynucleotides in a lane of a gel,staining the electrophoresed polynucleotides,forming an image of having an absorbance distribution related to the gel containing the stained polynucleotides,scanning the image of the gel lane to convert the electrophoresed polynucleotides into pixel-by-pixel representations of the absorbance Ai distribution of the electrophoresed polynucleotides, in terms of absorbance Ai and size Si,calculating the number average molecular size Sn of the polynucleotides distribution by the formula
calculating the weight average molecular size Sw of the polynucleotides distribution by the formula
Ai/Σ
Ai/Si
space="preserve" listing-type="equation">Sw=Σ
Ai Si/Σ
Aicalculating the Z average molecular size Sz of the polynucleotides distribution by the formula
space="preserve" listing-type="equation">Sz=Σ
Ai Si.sup.2 /Σ
Ai Sithereby to characterize the polynucleotide by the criteria number average molecular size, weight average molecular size, and Z average molecular size.
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Abstract
Polydispersed, double-stranded and single-stranded polynucleotides are characterized according to their molecular size distribution. The polynucleotides are electrophoresed, stained, imaged and the image scanned and converted to pixel-by-pixel representations of the image density and size. The number and weight average molecular size are calculated along with the Z average molecular size.
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Citations
11 Claims
- 1. A method of characterizing polydispersed, single-stranded or double-stranded polynucleotides according to their molecular size distribution comprising the steps of
converting the polynucleotides to single-stranded polynucleotides, electrophoresing the single-stranded polynucleotides in a lane of a gel, staining the electrophoresed polynucleotides, forming an image of having an absorbance distribution related to the gel containing the stained polynucleotides, scanning the image of the gel lane to convert the electrophoresed polynucleotides into pixel-by-pixel representations of the absorbance Ai distribution of the electrophoresed polynucleotides, in terms of absorbance Ai and size Si, calculating the number average molecular size Sn of the polynucleotides distribution by the formula - space="preserve" listing-type="equation">Sn=Σ
Ai/Σ
Ai/Si
calculating the weight average molecular size Sw of the polynucleotides distribution by the formula
space="preserve" listing-type="equation">Sw=Σ
Ai Si/Σ
Aicalculating the Z average molecular size Sz of the polynucleotides distribution by the formula
space="preserve" listing-type="equation">Sz=Σ
Ai Si.sup.2 /Σ
Ai Sithereby to characterize the polynucleotide by the criteria number average molecular size, weight average molecular size, and Z average molecular size. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11)
- 3. The method set forth in claim 2 which includes the additional step of calculating the skewness Z of the polynucleotide distribution by the formula
- space="preserve" listing-type="equation">Z=Sz/Sn.
- space="preserve" listing-type="equation">Sn=Σ
- space="preserve" listing-type="equation">Z=Sz/Sn.
- space="preserve" listing-type="equation">D=Sw/Sn; and
space="preserve" listing-type="equation">Z=Sz/Sn.
- space="preserve" listing-type="equation">D=Sw/Sn; and
space="preserve" listing-type="equation">Z=Sz/Sn.
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(a) summing the first/next eleven representations, (b) determining the means of those representations, (c) assign the mean as a first/next smoothed representation, (d) increment the series of representations by one, (e) repeat steps (a) through (d).
Specification