A whole-genome amplification protocol for a wide variety of DNAs, including those from formalin-fixed and paraffin-embedded tissue.
High-resolution genomic arrays and next-generation sequencers are some of the genome-based technologies poised to make significant contributions in the near future to basic and clinical science. The success of these technologies, and most certainly their translation into the clinic, will require that they produce high quality, reproducible data from small archived tumor specimens, including biopsies. DNA from patient samples, especially archival tissue, can be a limiting factor and lead to the need for amplification of the starting material. A variety of whole-genome amplification techniques are available, but choosing the most reliable, reproducible amplification technology that will be suitable for use across a wide spectrum of clinical specimens is essential. Sigma's whole-genome amplification kit provides a robust, highly reliable, and versatile amplification system across a variety of DNA sources. This chapter will detail Sigma's amplification protocol along with an optimized DNA extraction protocol for formalin-fixed and paraffin-embedded tissue.
Paris
Pamela L
PL
Department of Urology, University of California at San Francisco, San Francisco, CA, USA.
D055028:Comparative Genomic Hybridization; D004247:DNA / Q000737:chemistry / Q000235:genetics; D005557:Formaldehyde / Q000737:chemistry*; D016678:Genome; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation / Q000379:methods*; D016612:Paraffin Embedding; D011933:Reagent Kits, Diagnostic; D016707:Tissue Fixation / Q000379:methods*
D011933:Reagent Kits, Diagnostic; D005557:Formaldehyde; D004247:DNA