Table 2.

Pre-analytical recommendations for BRCA testing

Sample type	Key recommendations
Tissue sample	Follow College of American Pathologists guidelines for fixation of tissue samples (Hammond 2010), 8–48 hr, depending on size of specimen
	Vacuum and controlled temperature
	Controlled parafinization protocols
	Large samples must be properly handle by a pathologist or well-trained personnel in order to assure formaldehyde penetration and adequate fixation
	Cytological samples also useful because alcoholic fixatives excellently preserve DNA
	Sample must contain a percentage of tumor cells that is at least 30%. Avoid inflammation, immune infiltrates, and necrosis
	Morphological diagnosis should always include the type and grade of the tumors in order to gain insight into genotypic/phenotypic correlations
	Decalcification reduces DNA yield and quality. If needed, avoid acid decalcification
Blood sample	EDTA or nucleic acids preserving agent tubes
	Sample processed immediately after its extraction (ideally in less than 30 min when tubes contain no preservatives)
	Tubes treated with heparin must be avoided, since it can inhibit PCR reaction
	Centrifugation of the blood sample must be performed at 4 °C using low speed (1000–2000 g for 10 min), to split the hematic pellet from the fractions corresponding to the buffy coat and the plasma
	Higher centrifugation speed and time should be avoided to prevent platelet and leucocyte lysis
	Fractions obtained after blood centrifugation can be stored at –20 °C during short periods of time and at –80 °C for long terms
	Leucocyte fraction, serum, plasma, and whole blood are acceptable for germline DNA (leucocytes preferred)