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DNA Probe Labeling: Limited by low Amounts of gDNA Template?

WGA-based Labeling Reagents

Preparation of labeled DNA hybridization probes is often hampered by limited amounts of genomic DNA (gDNA) template. This drawback can be overcome by isothermal whole genome amplification (WGA) with Phi29 DNA polymerase whose strand displacement activity, high processivity and 3’-5’ exonuclease proofreading activity ensures efficient, minimal bias gDNA amplification.1-5

Labeled DNA hybridization probes from limited amount of purified gDNA can be produced in a four step procedure:


Selected References

[1] Dean et al. (2002) Comprehensive human genome amplification using multiple displacement amplification. Proc. Natl. Acad. Sci 99 (8):5261.
[2] Hosono et al. (2003) Unbiased whole-genome amplification directly from clinical samples. Genome Research 13:954.
[3] Hostetter et al. (2010) Random DNA fragmentation allows detection of single-copy, single-exon alterations of copy number by oligonucleotide array CGH in clinical FFPE samples. Nucleic Acid Res. 38 (2): e9.
[4] Kawachi et al. (2013) Deletion polymorphism at chromosome 3q26.1 and oral squamous cell carcinoma. International Journal of Oncology 42:384.
[5] Uda et al. (2007) Comparison of Whole Genome Amplification Methods for Detection of Pathogenic Bacterial Genomic DNA Using Microarray. Jpn. J. Infect. Dis. 60:355.