Exact(2)
At later stages of the process, the laboratory should also monitor the sample integrity, for example, for the presence of microbial degradation.
Samples with RNA integrity numbers less than or equal to 7 showed a significant increase in false positives relative to undegraded liver RNA and a reduction in the detection of true positives among probe sets most sensitive to sample integrity for in silico modeled changes of 1.5-, 2-, and 4-fold.
Similar(58)
Biobank sample integrity is paramount for subsequent studies but also other factors are important for showing the community that we are acting responsibly.
Successful dye incorporation and sample integrity was assessed for 1 μL aliquots of labeled samples by agarose gel electrophoresis followed by fluorescent detection of aRNA products (Typhoon scanner, GE Healthcare Life Sciences).
The accuracy of true positive detection of in silico modeled changes of 1.5-, 2-, and 4-fold was reduced for RNA samples with RIN values ≤ 7. The effect was greatest for probe sets most sensitive to sample integrity and was less pronounced for probe sets unselected for an effect of RNA degradation on signal level.
Total RNA samples were visually assessed on 1% (w/v) agarose gels for sample integrity before proceeding to cDNA synthesis.
It is essential to stress that swab samples should be obtained by vigorous sampling to acquire sufficient biologic material for testing, and development of a quality-control PCR target (housekeeping gene target) would be beneficial for sample integrity assessment, which is a limitation of this study.
The specificity of the PCR product was confirmed by sequencing with BigDyeTM terminator chemistry using the genetic analyser ABI Prism 310 (Applied Biosystems), β-2-microglobulin (as control for sample integrity) was amplified in the amplification mixture described above and using the following primers: 5'-TCTGGGTTTCATCCATCCGA-3' and 5'-CCCCAAATTCTAAGCAGAGTATGTAA-3'.
Sample integrity was maintained at 2 8 °C for the entire 30 d.
One adolescent was excluded from the final analysis due to his inconsistency in participating in the study activities and two other participants were excluded for questionable sample integrity, resulting in a final sample size of 95 (57 applicators and 38 non-applicators).
For verification of sample integrity, a 278 bp region of the pig mitochondrial NADH dehydrogenase subunit 1 (ND1) gene was amplified with primers JH0183 (5′-TCA TCG GGG CCC TAC GAG CA-3′) and JH0184 (5′-GGC GAA AGG TCC GGC TGC AT-3′).
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