Probes, libraries and kits for analysis of mixtures of nucleic acids and methods for constructing the same

Probes, libraries and kits for analysis of mixtures of nucleic acids and methods for constructing the same

  • CN 1,833,034 B
  • Filed: 06/18/2004
  • Issued: 04/16/2014
  • Est. Priority Date: 06/20/2003
  • Status: Active Grant
First Claim
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1. select can be used for the method for the oligonucleotide sequence in oligonucleotide probe library, compriseA) provide have predetermined Nucleotide number N likely oligonucleotide first set, wherein said oligonucleotide has the length of 8-9 Nucleotide and the melting temperature (Tm) T of at least 50 ℃

  • m, B) provide the second set of target nucleic acid sequence, described the second set is the people'"'"'s transcript group sequence obtaining from ENSMEBL,C) for each member of described the first set, from described the second set, identify respectively and storage comprises the number with the member of the sequence of this member'"'"'s complementation of described the first set,D) be chosen in the member of described first set of mating with the described second member who gathers of the maximum number identifying in step c in the evaluation of step c,E) member who selects in steps d is added into by select, can be used in the 3rd set that oligonucleotide in described oligonucleotide probe library forms,F) member who selects in deduction step d from described the first set to be so that the first set of revision to be provided,M) repeating step d to f, until the forming member of described the 3rd set combine can with target nucleic acid sequence set from step b at least 30% member'"'"'s complementation,1) wherein the target of each probe be present in sequence in described people'"'"'s transcript group at least 6% in,2) these oligonucleotide are not self complementations,3) the double-helical T that probe and its target sequence form mvalue is 60 ℃

    or higher, and described oligonucleotide covers the mRNA of >

    98% in human transcription thing group when combining, and 4) described library comprises the probe that the listed sequence of specification sheets table 1 obtains.

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