Gene expression studies are reported to be influenced by pre-analytical factors that can compromise RNA yield and integrity, which in turn may confound the experimental findings. Here we investigate the impact of four pre-analytical factors on brain-derived RNA: time-before-collection, tissue specimen size, tissue collection method, and RNA isolation method. We report no significant differences in RNA yield or integrity between 20 mg and 60 mg tissue samples collected in either liquid nitrogen or the RNAlater stabilizing solution. Isolation of RNA employing the TRIzol reagent resulted in a higher yield compared to isolation via the QIAcube kit while the latter resulted in RNA of slightly better integrity. Keeping brain tissue samples at room temperature for up to 160 min prior to collection and isolation of RNA resulted in no significant difference in yield or integrity. Our findings have significant practical and financial consequences for clinical genomic departments and other laboratory settings performing large-scale routine RNA expression analysis of brain samples.
机构:
Himalayan Inst Med Sci, Dept Transfus Med, Dehra Dun, Uttarakhand, India
Himalayan Inst Med Sci, Blood Bank, Dehra Dun, Uttarakhand, IndiaHimalayan Inst Med Sci, Dept Transfus Med, Dehra Dun, Uttarakhand, India
Jain, Ashish
Jain, Sanchit
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All India Inst Med Sci, Dept Forens Med, Rishikesh, Uttarakhand, IndiaHimalayan Inst Med Sci, Dept Transfus Med, Dehra Dun, Uttarakhand, India
Jain, Sanchit
Singh, Neha
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All India Inst Med Sci, Dept Pathol & Lab Med, Rishikesh 249203, Uttarakhand, IndiaHimalayan Inst Med Sci, Dept Transfus Med, Dehra Dun, Uttarakhand, India
Singh, Neha
Meinia, Sushant Kumar
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All India Inst Med Sci, Dept Transfus Med, Rishikesh, Uttarakhand, IndiaHimalayan Inst Med Sci, Dept Transfus Med, Dehra Dun, Uttarakhand, India
Meinia, Sushant Kumar
Chowdhury, Nilotpal
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All India Inst Med Sci, Dept Pathol & Lab Med, Rishikesh 249203, Uttarakhand, IndiaHimalayan Inst Med Sci, Dept Transfus Med, Dehra Dun, Uttarakhand, India
机构:
AstraZeneca, Personalised Healthcare & Biomarkers, Innovat Med & Early Dev Biotech Unit, Darwin Bldg,310 Cambridge Sci Pk,Milton Rd, Cambridge CB4 0WG, EnglandAstraZeneca, Personalised Healthcare & Biomarkers, Innovat Med & Early Dev Biotech Unit, Darwin Bldg,310 Cambridge Sci Pk,Milton Rd, Cambridge CB4 0WG, England
Sherwood, James L.
Corcoran, Claire
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AstraZeneca, Personalised Healthcare & Biomarkers, Innovat Med & Early Dev Biotech Unit, Darwin Bldg,310 Cambridge Sci Pk,Milton Rd, Cambridge CB4 0WG, EnglandAstraZeneca, Personalised Healthcare & Biomarkers, Innovat Med & Early Dev Biotech Unit, Darwin Bldg,310 Cambridge Sci Pk,Milton Rd, Cambridge CB4 0WG, England
Corcoran, Claire
Brown, Helen
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AstraZeneca, Personalised Healthcare & Biomarkers, Innovat Med & Early Dev Biotech Unit, Darwin Bldg,310 Cambridge Sci Pk,Milton Rd, Cambridge CB4 0WG, EnglandAstraZeneca, Personalised Healthcare & Biomarkers, Innovat Med & Early Dev Biotech Unit, Darwin Bldg,310 Cambridge Sci Pk,Milton Rd, Cambridge CB4 0WG, England
Brown, Helen
Sharpe, Alan D.
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AstraZeneca, Personalised Healthcare & Biomarkers, Innovat Med & Early Dev Biotech Unit, Darwin Bldg,310 Cambridge Sci Pk,Milton Rd, Cambridge CB4 0WG, EnglandAstraZeneca, Personalised Healthcare & Biomarkers, Innovat Med & Early Dev Biotech Unit, Darwin Bldg,310 Cambridge Sci Pk,Milton Rd, Cambridge CB4 0WG, England
Sharpe, Alan D.
Musilova, Milena
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AstraZeneca, Personalised Healthcare & Biomarkers, Innovat Med & Early Dev Biotech Unit, Darwin Bldg,310 Cambridge Sci Pk,Milton Rd, Cambridge CB4 0WG, EnglandAstraZeneca, Personalised Healthcare & Biomarkers, Innovat Med & Early Dev Biotech Unit, Darwin Bldg,310 Cambridge Sci Pk,Milton Rd, Cambridge CB4 0WG, England
Musilova, Milena
Kohlmann, Alexander
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AstraZeneca, Personalised Healthcare & Biomarkers, Innovat Med & Early Dev Biotech Unit, Darwin Bldg,310 Cambridge Sci Pk,Milton Rd, Cambridge CB4 0WG, EnglandAstraZeneca, Personalised Healthcare & Biomarkers, Innovat Med & Early Dev Biotech Unit, Darwin Bldg,310 Cambridge Sci Pk,Milton Rd, Cambridge CB4 0WG, England