Yield and Integrity of RNA from Brain Samples are Largely Unaffected by Pre-analytical Procedures
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Yield and Integrity of RNA from Brain Samples are Largely Unaffected by Pre-analytical Procedures. / Jensen, Pernille Søs Hovgaard; Johansen, Maja; Bak, Lasse K.; Jensen, Lars Juhl; Kjær, Christina.
In: Neurochemical Research, Vol. 46, 2021, p. 447-454.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Yield and Integrity of RNA from Brain Samples are Largely Unaffected by Pre-analytical Procedures
AU - Jensen, Pernille Søs Hovgaard
AU - Johansen, Maja
AU - Bak, Lasse K.
AU - Jensen, Lars Juhl
AU - Kjær, Christina
PY - 2021
Y1 - 2021
N2 - 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.
AB - 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.
KW - RNA isolation
KW - RNA Integrity Number (RIN)
KW - Brain tissue
KW - Pre-analytical factors
KW - Gene expression
KW - RNA stability
KW - RNA degradation
KW - GENE-EXPRESSION
KW - TISSUE
KW - QUALITY
KW - PRESERVATION
KW - STABILITY
KW - STABILIZATION
KW - MORPHOLOGY
KW - FROZEN
U2 - 10.1007/s11064-020-03183-z
DO - 10.1007/s11064-020-03183-z
M3 - Journal article
C2 - 33249516
VL - 46
SP - 447
EP - 454
JO - Neurochemical Research
JF - Neurochemical Research
SN - 0364-3190
ER -
ID: 253446035