A protocol for urine collection and storage prior to DNA methylation analysis.

ABSTRACT:

Background

Urine poses an attractive non-invasive means for obtaining liquid biopsies for oncological diagnostics. Especially molecular analysis on urinary DNA is a rapid growing field. However, optimal and practical storage conditions that result in preservation of urinary DNA, and in particular hypermethylated DNA (hmDNA), are yet to be determined.

Aim

To determine the most optimal and practical conditions for urine storage that result in adequate preservation of DNA for hmDNA analysis.

Methods

DNA yield for use in methylation analysis was determined by quantitative methylation specific PCR (qMSP) targeting the ACTB and RASSF1A genes on bisulfite modified DNA. First, DNA yield (ACTB qMSP) was determined in a pilot study on urine samples of healthy volunteers using t

SUBMITTER: Bosschieter J 

PROVIDER: S-EPMC6108475 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature