Biopolym. Cell. 2001; 17(3):253-255.
http://dx.doi.org/10.7124/bc.0005B5
Short Communications
Formamide and glycerol in sequencing of GC-rich DNA
1, 2Kvasha S. M., 2, 3, 4Zabarovsky E. R., 1, 2, 3Kashuba V. I., 1Rynditch A. V.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680
  2. Center for Genomics and Bioinformatics,
    Karolinska Institutet
    SE-171 77 Stockholm, Sweden
  3. Department of Microbiology, Tumor and Cell Biology,
    Karolinska Institute
    Nobels vag 5 171 65 Solna, Sweden
  4. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences
    32, Vavilova Str., Moscow, Russian Federation, 119991

Abstract

Formamide and glycerol are known to increase the sequencing efficiency. We compared the ability of different concentration of these co-solvents to improve the sequencing results of GC-rich template. Herein, we report that addition of 5 % formamide and 7.5–10 % glycerol is the most effective for sequencing GC-rich templates.
Full text: (PDF, in Russian)

References

[1] Kozak M. An analysis of vertebrate mRNA sequences: intimations of translational control. J Cell Biol. 1991;115(4):887-903.
[2] Hengen PN. Cycle sequencing through GC-rich regions. Trends Biochem Sci. 1996;21(1):33-4.
[3] Agarwal RK, Perl A. PCR amplification of highly GC-rich DNA template after denaturation by NaOH. Nucleic Acids Res. 1993;21(22):5283-4.
[4] Sun Y, Hegamyer G, Colburn NH. PCR-direct sequencing of a GC-rich region by inclusion of 10% DMSO: application to mouse c-jun. Biotechniques. 1993;15(3):372-4.
[5] Zhang W, Reading C, Deisseroth AB. Improved PCR sequencing with formamide. Trends Genet. 1992;8(10):332.
[6] Zhang W, Hu GY, Deisseroth A. Improvement of PCR sequencing by formamide. Nucleic Acids Res. 1991;19(23):6649.