Serum and Plasma Copy Number Detection Using Real-time PCR

Samanta Salvi; Vincenza Conteduca; Filippo Martignano; Giorgia Gurioli; Daniele Calistri; Valentina Casadio

doi:10.3791/56502

Journal

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Cancer Research

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Serum and Plasma Copy Number Detection Using Real-time PCR

JoVE Journal
Cancer Research

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JoVE Journal Cancer Research

Serum and Plasma Copy Number Detection Using Real-time PCR

11,454 Views

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09:21 min

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December 15, 2017

DOI:

10.3791/56502-v

DOI:

10.3791/56502-v

•

09:21 min

December 15, 2017

•

11434 Views

¹Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS

Transcript

The overall goal of this method is to detect the gene copy number variation in serum or plasma samples.This method can help answer key questions in advanced prostate cancer field such as detection of markers useful to predict or monitor the spores reduction.The main advantage of this technique is that it is easy and fast to do.The implications of this technique extend toward therapy of advanced prostate cancer because androgen receptor gene copy number variation is a frequent event.Generally, individuals new to this method will struggle because the concentration and the quality of cell-free DNA is often lower than expected.For serum collection, work from five milliliter samples of whole blood and serum without anticoagulants maintained at four degrees Celsius.Start by centrifuging the samples for 15 minutes at 1, 000 g.The supernatant is the serum.Transfer it into two-milliliter tubes and discard the pellet.Unless the serum is processed immediately, freeze the samples at 80

Summary

This manuscript describes the copy number variation analysis performed in serum or plasma DNA using real-time PCR approach. This method is suitable for the prediction of drug resistance in castration resistant prostate cancer patients, but it could be informative also for other diseases.