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Proc Natl Acad Sci U S A. Jul 1, 1992; 89(13): 5847–5851.
PMCID: PMC49394

Whole genome amplification from a single cell: implications for genetic analysis.

Abstract

We have developed an in vitro method for amplifying a large fraction of the DNA sequences present in a single haploid cell by repeated primer extensions using a mixture of 15-base random oligonucleotides. We studied 12 genetic loci and estimate that the probability of amplifying any sequence in the genome to a minimum of 30 copies is not less than 0.78 (95% confidence). Whole genome amplification beginning with a single cell, or other samples with very small amounts of DNA, has significant implications for multipoint mapping by sperm or oocyte typing and possibly for genetic disease diagnosis, forensics, and the analysis of ancient DNA samples.

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Selected References

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  • Saiki RK, Scharf S, Faloona F, Mullis KB, Horn GT, Erlich HA, Arnheim N. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science. 1985 Dec 20;230(4732):1350–1354. [PubMed]
  • Mullis KB, Faloona FA. Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. Methods Enzymol. 1987;155:335–350. [PubMed]
  • Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. [PubMed]
  • Li HH, Gyllensten UB, Cui XF, Saiki RK, Erlich HA, Arnheim N. Amplification and analysis of DNA sequences in single human sperm and diploid cells. Nature. 1988 Sep 29;335(6189):414–417. [PubMed]
  • Jeffreys AJ, Wilson V, Neumann R, Keyte J. Amplification of human minisatellites by the polymerase chain reaction: towards DNA fingerprinting of single cells. Nucleic Acids Res. 1988 Dec 9;16(23):10953–10971. [PMC free article] [PubMed]
  • Handyside AH, Pattinson JK, Penketh RJ, Delhanty JD, Winston RM, Tuddenham EG. Biopsy of human preimplantation embryos and sexing by DNA amplification. Lancet. 1989 Feb 18;1(8634):347–349. [PubMed]
  • Holding C, Monk M. Diagnosis of beta-thalassaemia by DNA amplification in single blastomeres from mouse preimplantation embryos. Lancet. 1989 Sep 2;2(8662):532–535. [PubMed]
  • Bradbury MW, Isola LM, Gordon JW. Enzymatic amplification of a Y chromosome repeat in a single blastomere allows identification of the sex of preimplantation mouse embryos. Proc Natl Acad Sci U S A. 1990 Jun;87(11):4053–4057. [PMC free article] [PubMed]
  • Gomez CM, Muggleton-Harris AL, Whittingham DG, Hood LE, Readhead C. Rapid preimplantation detection of mutant (shiverer) and normal alleles of the mouse myelin basic protein gene allowing selective implantation and birth of live young. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4481–4484. [PMC free article] [PubMed]
  • Monk M, Holding C. Amplification of a beta-haemoglobin sequence in individual human oocytes and polar bodies. Lancet. 1990 Apr 28;335(8696):985–988. [PubMed]
  • Strom CM, Verlinsky Y, Milayeva S, Evsikov S, Cieslak J, Lifchez A, Valle J, Moise J, Ginsberg N, Applebaum M. Preconception genetic diagnosis of cystic fibrosis. Lancet. 1990 Aug 4;336(8710):306–307. [PubMed]
  • Cui XF, Li HH, Goradia TM, Lange K, Kazazian HH, Jr, Galas D, Arnheim N. Single-sperm typing: determination of genetic distance between the G gamma-globin and parathyroid hormone loci by using the polymerase chain reaction and allele-specific oligomers. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9389–9393. [PMC free article] [PubMed]
  • Goradia TM, Stanton VP, Jr, Cui XF, Aburatani H, Li HH, Lange K, Housman DE, Arnheim N. Ordering three DNA polymorphisms on human chromosome 3 by sperm typing. Genomics. 1991 Jul;10(3):748–755. [PubMed]
  • Hubert R, Stanton VP, Jr, Aburatani H, Warren J, Li H, Housman DE, Arnheim N. Sperm typing allows accurate measurement of the recombination fraction between D3S2 and D3S3 on the short arm of human chromosome 3. Genomics. 1992 Apr;12(4):683–687. [PubMed]
  • Arnheim N, Li HH, Cui XF. PCR analysis of DNA sequences in single cells: single sperm gene mapping and genetic disease diagnosis. Genomics. 1990 Nov;8(3):415–419. [PubMed]
  • Li H, Cui X, Arnheim N. Direct electrophoretic detection of the allelic state of single DNA molecules in human sperm by using the polymerase chain reaction. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4580–4584. [PMC free article] [PubMed]
  • Lüdecke HJ, Senger G, Claussen U, Horsthemke B. Cloning defined regions of the human genome by microdissection of banded chromosomes and enzymatic amplification. Nature. 1989 Mar 23;338(6213):348–350. [PubMed]
  • Saunders RD, Glover DM, Ashburner M, Siden-Kiamos I, Louis C, Monastirioti M, Savakis C, Kafatos F. PCR amplification of DNA microdissected from a single polytene chromosome band: a comparison with conventional microcloning. Nucleic Acids Res. 1989 Nov 25;17(22):9027–9037. [PMC free article] [PubMed]
  • Kinzler KW, Vogelstein B. Whole genome PCR: application to the identification of sequences bound by gene regulatory proteins. Nucleic Acids Res. 1989 May 25;17(10):3645–3653. [PMC free article] [PubMed]
  • Lewin HA, Schmitt K, Hubert R, van Eijk MJ, Arnheim N. Close linkage between bovine prolactin and BoLA-DRB3 genes: genetic mapping in cattle by single sperm typing. Genomics. 1992 May;13(1):44–48. [PubMed]
  • Goradia TM, Lange K. Multilocus ordering strategies based on sperm typing. Ann Hum Genet. 1990 Jan;54(Pt 1):49–77. [PubMed]
  • Boehnke M, Arnheim N, Li H, Collins FS. Fine-structure genetic mapping of human chromosomes using the polymerase chain reaction on single sperm: experimental design considerations. Am J Hum Genet. 1989 Jul;45(1):21–32. [PMC free article] [PubMed]
  • Navidi W, Arnheim N. Using PCR in preimplantation genetic disease diagnosis. Hum Reprod. 1991 Jul;6(6):836–849. [PubMed]
  • Navidi W, Arnheim N, Waterman MS. A multiple-tubes approach for accurate genotyping of very small DNA samples by using PCR: statistical considerations. Am J Hum Genet. 1992 Feb;50(2):347–359. [PMC free article] [PubMed]

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