Item request has been placed! ×
Item request cannot be made. ×
loading  Processing Request

Single nucleotide polymorphism detection by combinatorial fluorescence energy transfer tags and biotinylated dideoxynucleotides.

Item request has been placed! ×
Item request cannot be made. ×
loading   Processing Request
اقرأ على الانترنت اقرأ أكثر حفظ في قائمتي
  • المؤلفون: Tong AK;Tong AK; Ju J
  • المصدر:
    Nucleic acids research [Nucleic Acids Res] 2002 Mar 01; Vol. 30 (5), pp. e19.
  • نوع النشر :
    Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.
  • اللغة:
    English
  • معلومة اضافية
    • المصدر:
      Publisher: Oxford University Press Country of Publication: England NLM ID: 0411011 Publication Model: Print Cited Medium: Internet ISSN: 1362-4962 (Electronic) Linking ISSN: 03051048 NLM ISO Abbreviation: Nucleic Acids Res Subsets: MEDLINE
    • بيانات النشر:
      Publication: 1992- : Oxford : Oxford University Press
      Original Publication: London, Information Retrieval ltd.
    • الموضوع:
      Polymorphism, Single Nucleotide*; DNA Primers/*chemistry ; Sequence Analysis, DNA/*methods; Biotinylation ; DNA-Directed DNA Polymerase/chemistry ; Dideoxynucleosides/chemistry ; Electrophoresis/methods ; Energy Transfer ; Fluorescence ; Fluorescent Dyes/chemistry ; Humans ; Polymerase Chain Reaction ; Retinoblastoma Protein/genetics
    • نبذة مختصرة :
      Combinatorial fluorescence energy transfer (CFET) tags, constructed by exploiting energy transfer and combinatorial synthesis, allow multiple biological targets to be analyzed simultaneously. We here describe a multiplex single nucleotide polymorphism (SNP) assay based on single base extension (SBE) using CFET tags and biotinylated dideoxynucleotides (biotin-ddNTPs). A library of CFET-labeled oligonucleotide primers was mixed with biotin-ddNTPs, DNA polymerase and the DNA templates containing the SNPs in a single tube. The nucleotide at the 3'-end of each CFET-labeled oligonucleotide primer was complementary to a particular SNP in the template. Only the CFET-labeled primer that is fully complementary to the DNA template was extended by DNA polymerase with a biotin-ddNTP. We isolated the DNA extension fragments that carry a biotin at the 3'-end by capture with streptavidin-coated magnetic beads, while the unextended primers were eliminated. The biotinylated fluorescent DNA fragments were subsequently analyzed in a multicolor fluorescence electrophoresis system. The distinct fluorescence signature and electrophoretic mobility of each DNA extension product in the electropherogram coded the SNPs without the use of a sizing standard. We simultaneously distinguished six nucleotide variations in synthetic DNA templates and a PCR product from the retinoblastoma tumor suppressor gene. The use of CFET-labeled primers and biotin-ddNTPs coupled with the specificity of DNA polymerase in SBE offered a multiplex method for detecting SNPs.
    • References:
      Nucleic Acids Res. 2001 Jul 15;29(14):E71. (PMID: 11452040)
      Genomics. 2001 Jun 15;74(3):402-7. (PMID: 11414769)
      Nucleic Acids Res. 2001 Nov 1;29(21):E104-4. (PMID: 11691941)
      Proc Natl Acad Sci U S A. 1987 Jul;84(14):4767-71. (PMID: 3474623)
      Science. 1988 Aug 26;241(4869):1077-80. (PMID: 3413476)
      Genomics. 1990 Dec;8(4):684-92. (PMID: 2276739)
      Am J Hum Genet. 1991 Oct;49(4):746-56. (PMID: 1897522)
      Nucleic Acids Res. 1994 Oct 25;22(21):4527-34. (PMID: 7526344)
      Genomics. 1994 Sep 1;23(1):138-44. (PMID: 7829062)
      Genomics. 1996 May 15;34(1):107-13. (PMID: 8661030)
      Clin Chem. 1997 Jan;43(1):40-4. (PMID: 8990220)
      Genome Res. 1997 Apr;7(4):378-88. (PMID: 9110177)
      Anal Chem. 1997 Jun 1;69(11):2181-6. (PMID: 9183181)
      Nucleic Acids Res. 1997 Aug 1;25(15):3102-9. (PMID: 9224611)
      Genome Res. 1998 Jan;8(1):69-80. (PMID: 9445489)
      Science. 1998 May 15;280(5366):1077-82. (PMID: 9582121)
      Genome Res. 1998 Aug;8(8):769-76. (PMID: 9724323)
      Clin Chem. 1999 Aug;45(8 Pt 1):1141-7. (PMID: 10430777)
      J Histochem Cytochem. 1999 Sep;47(9):1179-88. (PMID: 10449539)
      Anal Biochem. 1999 Sep 10;273(2):221-8. (PMID: 10469493)
      Hum Mutat. 1999;14(4):283-8. (PMID: 10502774)
      Nucleic Acids Res. 1996 Mar 15;24(6):1144-8. (PMID: 8604350)
      Proc Natl Acad Sci U S A. 1995 Jul 3;92(14):6339-43. (PMID: 7603992)
      Genomics. 1995 Sep 1;29(1):152-62. (PMID: 8530065)
      Nature. 2000 Jun 15;405(6788):857-65. (PMID: 10866212)
      Mol Biotechnol. 2000 Mar;14(3):223-33. (PMID: 10890013)
      Electrophoresis. 2000 Jul;21(12):2352-8. (PMID: 10939445)
      Nat Biotechnol. 2000 Oct;18(10):1096-100. (PMID: 11017050)
      Anal Chem. 2001 Jan 15;73(2):157-64. (PMID: 11199960)
      Nature. 2001 Feb 15;409(6822):928-33. (PMID: 11237013)
      Nat Biotechnol. 2001 Aug;19(8):756-9. (PMID: 11479569)
    • الرقم المعرف:
      0 (DNA Primers)
      0 (Dideoxynucleosides)
      0 (Fluorescent Dyes)
      0 (Retinoblastoma Protein)
      EC 2.7.7.7 (DNA-Directed DNA Polymerase)
    • الموضوع:
      Date Created: 20020228 Date Completed: 20020311 Latest Revision: 20190501
    • الموضوع:
      20250114
    • الرقم المعرف:
      PMC101255
    • الرقم المعرف:
      10.1093/nar/30.5.e19
    • الرقم المعرف:
      11861924