Meridian’s qPCR mix outperformed the competitor—delivering reliable library prep from just 5 ng of gDNA, with fewer failures, higher fidelity, and cleaner sequencing results.
Figure 3: Sequencing Results from Amplified DNA Fragments A) SEQUENCING PERFORMANCE
Raw_reads
Clean_reads
Map_rate (%)
Targeted_rate (%)
Uniformity (>0.1x)
41,044,434
40,966,416
99.8
99.8
85.8
Commercial tNGS Kit
36,644,668
36,578,418
99.9
99.8
87.8
35,911,938
35,851,470
99.9
99.8
86.6
15,056,954
14,970,350
99.9
99.96
87.3
Direct DNA qPCR Blood Mix (MDX122)
23,806,042
23,710,302
99.9
99.7
88.7
16,411,848
16,312,476
99.9
99.6
89.7
B) VARIANT DETECTION
Lyo-Ready Direct DNA qPCR Blood Mix (MDX122)
Targeted Gene
cds
pos
Theoretical
Commercial tNGS Kit
BRACA1
c.3548A>G
17:43091983
7.5%
8.0%
7.8%
BRACA1
c.2458A>G
17:43092919
7.5%
8.8%
8.6%
BRACA1
c.2612C>T
17:4309291
15.0%
15.0%
14.3%
BRACA1
c.4327C>T
17:43082434
32.5%
32.8%
33.0%
BRACA1
c.4900A>G
17:43071077
32.5%
7.5%
7.6%
BRACA1
c.4258G>T
13:32338613
10.0%
31.3%
31.7%
BRACA2
c.8021dup
13:32363217
32.5%
8.3%
8.4%
BRACA2
c.5073del
13:32339421
40.0%
32.1%
32.6%
BRACA2
c.5351del
13:32339699
7.5%
37.0%
37.5%
BRACA2
c.86SA>C
13:32332343
7.5%
7.2%
7.3%
BRACA2
c.2971A>G
13:32337326
7.5%
6.6%
7.0%
BRACA2
c.7397T>C
13:3235525
100.0%
100.0%
99.6%
HORIZON BRCA Somatic Multiplex l gDNA Multiplex
Taken together, these findings confirm that Meridian’s Lyo-Ready Direct DNA qPCR Blood Mix (MDX122) effectively amplifies targeted DNA with high specificity and yield, producing high-quality sequencing libraries even from challenging sample types. The mix ensures reliable variant detection, particularly for low-frequency mutations, without compromising performance in a tNGS workflow. Figure 3. A) Sequencing reads of the commercial tNGS kit vs Meridian’s Lyo-Ready Direct DNA qPCR Blood Mix (MDX122); B) Variant detection in both groups. Both mixes show similar sequencing metrics with high mapping rates (>99%) and uniformity (>85%), reflecting consistent data quality. The observed allele frequencies in both mixes closely align with theoretical values for all variants, demonstrating accurate variant detection across experiments. Raw Reads (Raw_reads): Total raw reads generated from sequencing. Clean Reads (Clean_reads): Reads retained after filtering for quality. Mapping Rate (Map_rate%): Percentage of reads successfully mapped to the reference genome. Target Rate (Target_rate%): Percentage of reads aligned to the target regions. Uniformity (>0.1x): Measure of how evenly sequencing reads cover the target regions. CDS (Coding Sequence): Variants in the coding sequence. POS (Position): Genomic position of each variant. Theoretical: Expected allele frequency of the variant. Commercial tNGS Kit: Measured allele frequency in the control group. Lyo-Ready Direct DNA qPCR Mix (MDX122): Measured allele frequency in the experimental group. These findings validate that Meridian’s Lyo-Ready Direct DNA qPCR Blood (MDX122) provides precise and consistent amplification, supporting high-confidence variant mix detection in a targeted sequencing approach.
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