Low-cost, high-sensitivity laser-induced fluorescence detection for DNA sequencing by capillary gel electrophoresis - PubMed

Low-cost, high-sensitivity laser-induced fluorescence detection for DNA sequencing by capillary gel electrophoresis

D Y Chen et al. J Chromatogr. 1991.

Abstract

A low cost, 0.75-mW helium neon laser, operating in the green region at 534.5 nm, is used to excite fluorescence from tetramethylrhodamine isothiocyanate-labelled DNA fragments that have been separated by capillary gel electrophoresis. The detection limit (3 sigma) for the dye is 500 ymol [1 yoctomole (1 ymol) = 10(-24) mol] or 300 analyte molecules in capillary zone electrophoresis; the detection limit for labeled primer separated by capillary gel electrophoresis is 2 zmol [1 zeptomole (1 zmol) = 10(-21) mol]. The Richardson-Tabor peak-height encoded sequencing technique is used to prepare DNA sequencing samples. In 6% T, 5% C acrylamide, 7 M urea gels, sequencing rates of 300 bases/hour are produced at an electric field strength of 200 V/cm; unfortunately, the data are plagued by compressions. These compressions are eliminated with addition of 20% formamide to the sequencing gel; the gel runs slowly and sequencing data are generated at a rate of about 70 bases/hour.

Similar articles

• Two-label peak-height encoded DNA sequencing by capillary gel electrophoresis: three examples.

  Chen D, Harke HR, Dovichi NJ. Chen D, et al. Nucleic Acids Res. 1992 Sep 25;20(18):4873-80. doi: 10.1093/nar/20.18.4873. Nucleic Acids Res. 1992. PMID: 1408803 Free PMC article.

• Three DNA sequencing methods using capillary gel electrophoresis and laser-induced fluorescence.

  Swerdlow H, Zhang JZ, Chen DY, Harke HR, Grey R, Wu SL, Dovichi NJ, Fuller C. Swerdlow H, et al. Anal Chem. 1991 Dec 15;63(24):2835-41. doi: 10.1021/ac00024a006. Anal Chem. 1991. PMID: 1789449

• Capillary gel electrophoresis for DNA sequencing. Laser-induced fluorescence detection with the sheath flow cuvette.

  Swerdlow H, Wu SL, Harke H, Dovichi NJ. Swerdlow H, et al. J Chromatogr. 1990 Sep 7;516(1):61-7. doi: 10.1016/s0021-9673(01)90204-3. J Chromatogr. 1990. PMID: 2286629

• High-speed automated DNA sequencing in ultrathin slab gels.

  Smith LM, Brumley RL Jr, Buxton EC, Giddings M, Marchbanks M, Tong X. Smith LM, et al. Methods Enzymol. 1996;271:219-37. doi: 10.1016/s0076-6879(96)71012-1. Methods Enzymol. 1996. PMID: 8782556 Review. No abstract available.

• Separation and analysis of DNA by electrophoresis.

  Smith CL. Smith CL. Curr Opin Biotechnol. 1991 Feb;2(1):86-91. doi: 10.1016/0958-1669(91)90065-d. Curr Opin Biotechnol. 1991. PMID: 1367713 Review.

Cited by

• Electromigration behavior of polysaccharides in capillary electrophoresis under pulsed-field conditions.

  Sudor J, Novotny M. Sudor J, et al. Proc Natl Acad Sci U S A. 1993 Oct 15;90(20):9451-5. doi: 10.1073/pnas.90.20.9451. Proc Natl Acad Sci U S A. 1993. PMID: 8415721 Free PMC article.

• Reproducible two-dimensional capillary electrophoresis analysis of Barrett's esophagus tissues.

  Kraly JR, Jones MR, Gomez DG, Dickerson JA, Harwood MM, Eggertson M, Paulson TG, Sanchez CA, Odze R, Feng Z, Reid BJ, Dovichi NJ. Kraly JR, et al. Anal Chem. 2006 Sep 1;78(17):5977-86. doi: 10.1021/ac061029+. Anal Chem. 2006. PMID: 16944874 Free PMC article.

• Two-label peak-height encoded DNA sequencing by capillary gel electrophoresis: three examples.

  Chen D, Harke HR, Dovichi NJ. Chen D, et al. Nucleic Acids Res. 1992 Sep 25;20(18):4873-80. doi: 10.1093/nar/20.18.4873. Nucleic Acids Res. 1992. PMID: 1408803 Free PMC article.

• Effects of Repeated Ropinirole Treatment on Phencyclidine-Induced Hyperlocomotion, Prepulse Inhibition Deficits, and Social Avoidance in Rats.

  Maple AM, Call T, Kimmel PC, Hammer RP Jr. Maple AM, et al. J Pharmacol Exp Ther. 2017 Apr;361(1):109-114. doi: 10.1124/jpet.116.238634. Epub 2017 Feb 6. J Pharmacol Exp Ther. 2017. PMID: 28167638 Free PMC article.

Publication types

MeSH terms

Substances

LinkOut - more resources

• Other Literature Sources

  • The Lens - Patent Citations Database