hello Danielle Lewis It depends! If you are seeking biomarker discovery without prior knowledge you start with NGS. NGS is a high throughput analytical technique that utilizes bioinformatics to arrive at meaningful results. NGS is thus prone to human and technical error. QOCR is a simpler, direct analytical technique (not without potential errors, but is easier managed) and is used to confirm NGS findings. Though is recent years NGS has become somewhat reliable that many do not seek confirmation/validation by qPCR. So it is NGS then qPCR.

Hello Christina if you meant ngs count = the number of reads overlapping a given genetic region, it depends on the coverage and depth of your NGS data. If your coverage is high for these regions then higher than 20 counts should be expected, But it really depends on rarity of allele, type of sample (admixed or biopsy or fluid extract) and on input number of cells/ DNA. Lower counts are acceptable for degraded or FFPE samples, and low coverage experiments. In general, I set counts to be acceptable for 50x exome coverage after looking at overall data quality and experiment parameters for example, 30 for WBC DNA, and lower for cancer FFPE DNA. As i don't know what are your experimental parameters, I can't judge your 20 count threshold. There are no preset standards for NGS yet since there are many variables to consider. Consult (Article Standardization of Sequencing Coverage Depth in NGS: Recomme...