Applied study on magnetic nanometer beads in preparation of genechip samples

Abstract

A protocol for enrichment and adsorption of karyocyte from whole blood by using magnetic nanometer beads as solid-phase absorbents was presented. The PCR amplification could be accomplished by using the nanobeads with karyocyte as template directly and the PCR products were applied on an oligonucleotide array to do gene typing. The HLA-A PCR amplification system and a small HLA-A oligonucleotide microarray were applied as the platform and an experiment protocol of separating karyocyte from whole blood using the magnetic nanometer beads (Fe₂O₃) were set up. The experimental conditions were also discussed. It showed that pH level of PBS eluent, Taq enzyme quantity and fragment length of products could influent the amplification results, and the magnetic nano-beads could succeed in sample preparation in microarray to provide a promising way in automatic detection and lab-on-a-chip.

Keywords

sample preparation / magnetic nanometer bead / lab-on-a-chip

References

[1] RudiK, KrokenM, DahlbergO J, et al.. Rapid, universal method to isolate PCR-ready DNA using magnetic beads. Biotechniques, 1997, 22: 506-506

[2] DeggerdalA, LarsenF. Rapid isolation of PCR-ready DNA from blood, bone marrow and cultured cells, based on paramagnetic beads. Biotechniques, 1997, 22: 554-554

[3] DavisM J, TaylorJ I, SachsingerN, et al.. Isolation of plasmid DNA using magnetite as a solid-phase absorbent. Anal Biochem, 1998, 262: 92-92

[4] GaoX, JakobsenI B, SerjeantsonS W. Characterization of the HLA-A polymorphism by locus-specific polymerase chain reaction amplification and oligonucleotide hybridization. Hum Immunol, 1994, 41: 267-267

[5] Petzl-ErlerM L, McDevittH O. Molecular analysis of the HLA DRB genes in two tribes of Brazilian Indians. Hum Immunol, 1994, 41: 180-180