Top Abstract Purchase Related
Home
>
Node
>
Subnode

Dual asymmetric PCR: one-step construction of synthetic genes

Sandhu, G.S.; Aleff, R.A.; Kline, B.C.

Biotechniques 12(1): 14-16

1992

ISSN/ISBN: 0736-6205
PMID: 1734918
Document Number: 397523
We have developed a one-step process for constructing synthetic genes. Four adjacent oligonucleotides 17-100 bases in length having short overlaps of 15-17 bases are used as primers in a PCR mixture. The quantity of the two internal primers is highly limited, and the resultant reaction causes an asymmetric single-stranded amplification of the two halves of the total sequence due to an excess of the two flanking primers. In subsequent PCR cycles, these dual asymmetrically amplified fragments, which overlap each other, yield a double-stranded, full-length product.

Document emailed within 1 workday
Secure & encrypted payments

Van Dongen, J.J. 1988: Immunoglobulin genes and T-cell receptor genes and the expression of immunological markers. I. the construction of the specific immune system Nederlands Tijdschrift Voor Geneeskunde 132(19): 855-862
Bothwell, A.L.; Paskind, M.; Schwartz, R.C.; Sonenshein, G.E.; Gefter, M.L.; Baltimore, D. 1981: Dual expression of lambda genes in the MOPC-315 plasmacytoma Nature 290(5801): 65-67
Mitrunen, K.; Pettersson, K.; Piironen, T.; Björk, T.; Lilja, H.; Lövgren, T. 1995: Dual-label one-step immunoassay for simultaneous measurement of free and total prostate-specific antigen concentrations and ratios in serum Clinical Chemistry 41(8 Pt 1): 1115-1120
Nguyen, Q.; Witney, F.; Tumolo, A. 1992: A two-step hybridization method for chemiluminescent detection of single copy genes Biotechniques 13(1): 116-123
Qin, S.; Tang, H.; Zhao, L.; Liu, L.; Lin, Y. 2001: Construction of recombinant plasmids containing genes of HBsAg and their expression in the eukaryotic cells Hua Xi Yi Ke da Xue Xue Bao 32(2): 172-174
Douard, R.; Ettorre, G.Maria.; Sommacale, D.; Jan, D.; Révillon, Y.; Farges, O.; Belghiti, J. 2002: A two-step strategy for enlargement of left arterial branch in a living related liver graft with dual arterial supply Transplantation 73(6): 993-994
Nagata, Y.; Shiku, H. 1990: Dot hybridization with synthetic oligonucleotide; analysis of point mutation of c-ras genes Nihon Rinsho. Japanese Journal of Clinical Medicine 48(8): 163-169
Stekhin, I.N.; Shestopalova, N.S.; Panchenko, L.P.; Malinovskaia, L.P.; Skripal', I.G. 1989: The identification of restriction fragments containing rRNA genes and construction of a gene bank of Acholeplasma laidlawii PG8 Mikrobiolohichnyi Zhurnal 50(5): 41-45
Pan'kova, N.V.; Karamyshev, A.L.; Shliapnikov, M.G.; Presich, A.N.; Ksenzenko, V.N. 1998: Amber-suppressive tRNA from bacteriophage T5: construction of genes and determination of the effectiveness of suppression in vivo Molekuliarnaia Biologiia 32(6): 1004-1012
Kiselev, V.I.; Dubeĭkovskiĭ, A.N.; Pachkunov, D.M.; Astashkin, E.I.; Rechinskiĭ, V.O. 1987: Directed mutagenesis of chromosomal genes of Escherichia coli in vivo: construction of RecA- and HtpR- mutants Molekuliarnaia Genetika Mikrobiologiia i Virusologiia 4: 43-46
Yang, W.-t.; Liu, X.-m.; Wan, Q.; Yao, N.; Wang, N.; Zhang, X.-m.; Jiao, Z.-d.; Li, H.-y.; Li, X.-k. 2015: Full-length cDNA cloning of flavonol synthase genes of Carthamus tinctorius and construction plant expression vector Zhongguo Zhong Yao Za Zhi 40(4): 634-638
Pala, P.; Corradin, G.; Strachan, T.; Sodoyer, R.; Jordan, B.R.; Cerottini, J.C.; Maryanski, J.L. 1988: Mapping of HLA epitopes recognized by H-2-restricted cytotoxic T lymphocytes specific for HLA using recombinant genes and synthetic peptides Journal of Immunology 140(3): 871-877
Toyoda, H.; Onohara-Toyoda, M.; Krull, J.; Vadheim, C.M.; Bickal, J.C.; Riley, W.J.; MacLaren, N.K.; Itakura, K.; Rotter, J.I. 1989: Application of synthetic oligonucleotides to detect DQ beta genes transmission within insulin-dependent diabetes families Disease Markers 7(4): 215-228
Oliĭnyk, O.S.; Kaberniuk, A.A.; Redchuk, T.A.; Korotkevych, N.V.; Labyntsev, A.Iu.; Romaniuk, S.I.; Kolybo, D.V.; Komisarenko, S.V. 2009: Construction of immune library of murine immunoglobulin genes and screening of single-chain Fv-antibodies specific to diphtheria toxin B subunit Ukrains'kyi Biokhimichnyi Zhurnal 81(2): 68-79
Turetskaya, R.L.; Udalova, I.A.; Kuprash, D.V.; Terskikh, A.V.; Nedopasov, S.A. 1991: Use of synthetic oligonucleotides for studying gene structure and polymorphism: human genes encoding tumor necrosis factors and their receptors Nucleic Acids Symposium Series 24: 275
Ma, X-Li.; Wang, B-Bei.; Wu, P.; Lu, Y-Ping.; Zhou, J-Feng.; Ma, D. 2009: Construction of anti-sense cDNA library of human breast cancer cells during apoptosis induced by trichostatin A and preliminary screening of essential genes Zhonghua Yi Xue Za Zhi 89(7): 480-484
Paddock, G.V. 1989: Construction of mRNA genes for the synthesis and translation of duck alpha globin mRNA Biotechniques 7(8): 856-865
Song, X.; Liu, X.; Cai, L.; Wu, J.; Wang, J. 2015: Construction of cTnC-linker-Tni (P) Genes, Expression of Fusion Protein and Preparation of Lyophilized Protein Sheng Wu Yi Xue Gong Cheng Xue Za Zhi 32(6): 1267-1272
Chebotar, G.A.; Chebotar, S.V.; Motsnyy, I.I. 2016: Pleiotropic effects of Gibberellin-Sensitive and Gibberellin-Insensitive Dwarfing Genes in Common Wheat of the Southern Step Region of Black Sea Tsitologiia i Genetika 50(1): 26-35
Paul, R.A. 2016: Is the nature of psychoanalytic thinking and practice (e.g., in regard to sexuality) determined by extra-analytic, social and cultural developments?: Sexuality: Biological fact or cultural construction? The view from dual inheritance theory International journal of psycho-analysis 97(3): 823-837