Spectral calibration of fluorescent polynucleotide separation apparatus

Abstract

The invention relates to methods, compositions, and systems for calibrating a fluorescent polynucleotide separation apparatus.

1 .- 12 . (canceled)  
     
     
         13 . A system for calibrating a fluorescent polynucleotide separation apparatus, said system comprising a processor and, a computer readable medium functionally coupled to said processor for storing a computer program comprising: 
 computer code that receives plurality of spectral and temporal data from a fluorescent polynucleotide separation apparatus, and    computer code that calculates a total emission temporal profile from the spectral and temporal data.    
     
     
         14 . A calibration standard for a fluorescent polynucleotide separation apparatus, standard comprising four polynucleotides of different length, each polynucleotide labeled with a different fluorescent dye having a distinctive spectral profile having a peak, wherein the length of each of the polynucleotides is such that the peak of the spectral profile of each dye does not significantly overlap between the separated fragment.  
     
     
         15 . A kit for producing a calibration standard of  claim 14 , wherein the fluorescent labeled polynucleotides are stored in separate containers.  
     
     
         16 . A method of detecting the flow of electrical current through a separation channel of fluorescent polynucleotide separation apparatus, said method comprising the steps, introducing a fluorescently labeled polynucleotide composition to a channel of a fluorescent polynucleotide separation apparatus, said composition comprising 
 a polynucleotide labeled with a first florescent dye,    a monitoring dye that is spectrally distinct from the florescent dye, and    detecting the monitoring dye.    
     
     
         17 . The method according to  claim 16 , wherein the composition comprises a plurality of polynucleotides labeled with at least two spectrally distinct fluorescent dyes, wherein the monitoring dye is spectrally distinct from the florescent dyes.  
     
     
         18 . The method of  claim 17 , wherein the polynucleotides labeled with at least two spectrally distinct florescent dyes is a polynucleotide sequencing reaction product mixture.  
     
     
         19 . The method of  claim 18 , wherein the monitoring dye is attached to a polynucleotide.  
     
     
         20 . A composition for monitoring the flow of electric current through a separation channel of fluorescent polynucleotide separation apparatus, said composition comprising 
 a fluorescent dye labeled polynucleotide composition, and    a monitoring dye that is spectrally distinct from the florescent dyes of the polynucleotide composition.    
     
     
         21 . The composition according to  claim 20 , wherein the fluorescent dye labeled polynucleotide composition is a polynucleotide sequencing reaction product mixture.