Band array display of polynucleotide separations
First Claim
1. A method for producing an array of bands, each band representing a specific fraction of double stranded DNA having a calculated base-pair length, comprising the steps ofa) separating a mixture of double stranded nucleic acid fragments into base-pair length fractions by reversed phase ion pairing chromatography, wherein the separated base pair length fractions are eluted from a chromatography column;
- b) detecting the separated base-pair length fractions as they elute from the column;
c) providing a digitized signal corresponding to the detected base-pair length fractions; and
d) displaying the digitized signal as an array of bands, each band corresponding to a base-pair length fraction.
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Abstract
A method and apparatus for representing double stranded nucleic acid fragments which have been separated by a chromatographic process as an array of bands which can be accurately quantified, optimized and stored. Using, for example, a Matched Ion Polynucleotide Chromatography (MIPC) process, an analog output from a UV detector is digitized and input to a computer. The digitized signal is converted to a linear array of bands which may be displayed on a video display terminal. The intensity and/or color of a band may correlate to the amount of double stranded nucleic acid in the respective fraction or the respective double stranded nucleic acid fragment above a user selected threshold level at a corresponding point in the digitized signal. The calculated base pair length, concentration, and retention time of each band in the array of bands may be displayed in alphanumeric form.
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Citations
16 Claims
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1. A method for producing an array of bands, each band representing a specific fraction of double stranded DNA having a calculated base-pair length, comprising the steps of
a) separating a mixture of double stranded nucleic acid fragments into base-pair length fractions by reversed phase ion pairing chromatography, wherein the separated base pair length fractions are eluted from a chromatography column; -
b) detecting the separated base-pair length fractions as they elute from the column;
c) providing a digitized signal corresponding to the detected base-pair length fractions; and
d) displaying the digitized signal as an array of bands, each band corresponding to a base-pair length fraction. - View Dependent Claims (2, 3, 4, 5, 6, 7)
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8. An apparatus for producing an array of bands, each band representing a specific fraction of double stranded DNA having a calculated base-pair length, comprising:
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a) a reversed phase ion pairing chromatography system for separating a mixture of double stranded nucleic acid fragments into base-pair length fractions, wherein the chromatography system includes chromatography column with an outlet at one end from which the separated base-pair length fractions are eluted;
b) a detector positioned at the output of the column to detect the separated base-pair length fractions as they elute off the column; and
c) a computer and software to provide a digitized signal corresponding to the detected base-pair length fractions and to display the digitized signal as an array of bands, each band corresponding to a base-pair length fraction. - View Dependent Claims (9, 10, 11, 12, 13, 14, 15, 16)
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Specification