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QRT-PCR: Difference between revisions
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→Reagent lots: probe age
(→Sources of variability: lot variation) |
(→Reagent lots: probe age) |
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* person C: 2·7 × 10<sup>3</sup> different by a factor of 300!! | * person C: 2·7 × 10<sup>3</sup> different by a factor of 300!! | ||
=== Reagent lots === | === Reagent lots/age === | ||
Different lots of reagents can lead to different results. Experiment repeated by same operator 5 times, same RNA sample, different kits; values are copies/μg total RNA | Different lots of reagents can lead to different results. Experiment repeated by same operator 5 times, same RNA sample, different kits; values are copies/μg total RNA: | ||
* kit 1: 13±32 × 10<sup>7</sup> | * kit 1: 13±32 × 10<sup>7</sup> | ||
* kit 2: 5.4±1.6 × 10<sup>7</sup> - different by a factor of 2.4 | * kit 2: 5.4±1.6 × 10<sup>7</sup> - different by a factor of 2.4 | ||
Similar experiment with old (9 months 4°C) and new probe (3 months 4°C), values are copies/μg total RNA: | |||
* old: (5.6 ± 1.3) x 10<sup>3</sup> | |||
* new: (3.8 ± 0.6) x 10<sup>8</sup> - different by a factor of 100'000!! | |||
both experiments above from [Bustin 2002 PMID 12200227, figure 4] | |||
== Notes == | == Notes == | ||
