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Quant it broad range rna assay kit

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The Quant-iT Broad Range RNA Assay kit is a nucleic acid quantitation kit designed to measure the concentration of RNA in solution. The kit provides a sensitive and accurate method for determining the amount of RNA present in a sample.

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Lab products found in correlation

Agilent 2100 bioanalyzer, by Agilent Technologies (7 mentions) Mirvana paris kit, by Thermo Fisher Scientific (7 mentions) Standard sensitivity rna analysis dnf 471 kit, by Agilent Technologies (4 mentions) Qubit 3.0 fluorometer, by Thermo Fisher Scientific (3 mentions) 96 channel fragment analyzer, by Agilent Technologies (3 mentions) Rnalater, by Thermo Fisher Scientific (2 mentions) 48 channel fragment analyzer, by Agilent Technologies (2 mentions) Rlt buffer, by Qiagen (2 mentions) Rneasy 96 kit, by Qiagen (2 mentions) Qubit 2.0 fluorometer, by Thermo Fisher Scientific (1 mentions) Quantitect reverse transcription kit, by Qiagen (1 mentions) Tapestation rna screentape, by Agilent Technologies (1 mentions) Ncounter mouse myeloid innate immunity v2 panel, by NanoString (1 mentions) Zr bashingbead lysis tube, by Zymo Research (1 mentions) Geno grinder, by SPEX (1 mentions) Agilent tapestation rna screentape, by Agilent Technologies (1 mentions) Rsc simply rna tissue kit, by Promega (1 mentions) Maxwell 16 lev plant rna kit, by Promega (1 mentions) Tissuelyser 2, by Qiagen (1 mentions) Agilent rna 6000 nano kit, by Agilent Technologies (1 mentions)

17 protocols using quant it broad range rna assay kit

1

Nucleus Accumbens RNA Isolation

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Total RNA containing the small RNA fraction was isolated from 100mg of frozen tissue from the nucleus accumbens (NAc) using the mirVana-PARIS kit (Life Technologies, Carlsbad, CA), following manufacturer's protocols. RNA concentration was measured using the Quant-iT Broad Range RNA Assay kit (Life Technologies), and the RNA Integrity Number (RIN) was measured on the Agilent 2100 Bioanalyzer (Agilent Technologies, Inc., Santa Clara, CA). All subjects were initially included for matching based on age, sex, ethnicity, brain pH, PMI and RIN, and this yielded 18 appropriately matched case-control pairs with RINs ≥6 (N = 36).
Mamdani M., Williamson V., McMichael G.O., Blevins T., Aliev F., Adkins A., Hack L., Bigdeli T., D. van der Vaart A., Web B.T., Bacanu S.A., Kalsi G., Kendler K.S., Miles M.F., Dick D., Riley B.P., Dumur C, & Vladimirov V.I. (2015). Integrating mRNA and miRNA Weighted Gene Co-Expression Networks with eQTLs in the Nucleus Accumbens of Subjects with Alcohol Dependence. PLoS ONE, 10(9), e0137671.
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2

RNA Isolation from Whole Blood

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Total RNA containing the small RNA fraction was isolated from 9 ml of whole blood using the mirVana-PARIS kit (Life Technologies, Carlsbad, CA), following manufacturer's protocols. RNA concentration was measured using the Quant-iT Broad Range RNA Assay kit (Life Technologies), and the RNA Integrity Number (RIN) was measured on the Agilent 2100 Bioanalyzer (Agilent Technologies, Inc., Santa Clara, CA). All RNA samples had an excellent RIN scores (average RIN ≥ 9.0, s.d. ± 0.5).
Bountress K.E., Vladimirov V., McMichael G., Taylor Z.N., Hardiman G., Chung D., Adams Z.W., Danielson C.K, & Amstadter A.B. (2021). Gene Expression Differences Between Young Adults Based on Trauma History and Post-traumatic Stress Disorder. Frontiers in Psychiatry, 12, 581093.
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3

Total RNA Extraction from Frozen NAc

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Total RNA was isolated from 50 mg of frozen tissue from NAc using the mirVana‐PARIS kit (Thermo Fisher, Carlsbad, CA), following manufacturer's protocols. RNA concentration was measured using the Quant‐iT Broad Range RNA Assay kit (Life Technologies), and the RNA integrity number (RIN) was measured on the Agilent 2100 Bioanalyzer (Agilent Technologies, Inc., Santa Clara, CA).
Drake J., McMichael G.O., Vornholt E.S., Cresswell K., Williamson V., Chatzinakos C., Mamdani M., Hariharan S., Kendler K.S., Kalsi G., Riley B.P., Dozmorov M., Miles M.F., Bacanu S, & Vladimirov V.I. (2020). Assessing the Role of Long Noncoding RNA in Nucleus Accumbens in Subjects With Alcohol Dependence. Alcoholism, Clinical and Experimental Research, 44(12), 2468-2480.
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4

Postmortem NAc Transcriptomics of AD

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Postmortem NAc from 42 AD cases and 42 controls was provided by the Australian Brain Donor Programs of New South Wales Tissue Resource Centre (NSW TRC) under the support of the University of Sydney, National Health and Medical Research Council of Australia, Schizophrenia Research Institute, National Institute on Alcohol Abuse and Alcoholism, and the New South Wales Department of Health.26 (link) As part of a previous study,27 (link) several criteria were used to exclude samples with (1) agonal state, (2) substantial brain damage, (3) history of infectious disease, and (4) postmortem interval (PMI) > 48 h (Table S1). Samples were further matched for RNA integrity number (RIN) (mean = 6.9, ±0.84), sex (all male), ethnicity (100% Caucasian), brain pH, and postmortem interval to minimize covariates' effect on expression, resulting in 18 matched case–control pairs (n = 36).28 (link) Total RNA from flash-frozen NAc was extracted and purified via mirVANA-PARIS kit (Life Technologies, Carlsbad, CA) following the manufacturer's protocol. RNA integrity (RIN) and concentrations were assessed via Agilent 2100 Bioanalyzer (Agilent Technologies, Inc., Santa Clara, CA) and Quant-iT Broad Range RNA Assay kit (Life Technologies), respectively.
Vornholt E., Drake J., Mamdani M., McMichael G., Taylor Z.N., Bacanu S., Miles M.F, & Vladimirov V.I. (2021). Identifying a novel biological mechanism for alcohol addiction associated with circRNA networks acting as potential miRNA sponges. Addiction Biology, 26(6), e13071.
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5

Postmortem Analysis of Nucleus Accumbens

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Postmortem NAc from 42 AD cases and 42 controls was provided by the Australian Brain Donor Programs of New South Wales Tissue Resource Centre (NSW TRC) under the support of the University of Sydney, National Health and Medical Research Council of Australia, Schizophrenia Research Institute, National Institute on Alcohol Abuse and Alcoholism, and the New South Wales Department of Health.26 As part of a previous study,27 several criteria were used to exclude samples with (1) agonal state, (2) substantial brain damage, (3) history of infectious disease, and (4) postmortem interval (PMI) > 48 h (Table S1). Samples were further matched for RNA integrity number (RIN) (mean = 6.9, ±0.84), sex (all male), ethnicity (100% Caucasian), brain pH, and postmortem interval to minimize covariates' effect on expression, resulting in 18 matched case–control pairs (n = 36).28 Total RNA from flash‐frozen NAc was extracted and purified via mirVANA‐PARIS kit (Life Technologies, Carlsbad, CA) following the manufacturer's protocol. RNA integrity (RIN) and concentrations were assessed via Agilent 2100 Bioanalyzer (Agilent Technologies, Inc., Santa Clara, CA) and Quant‐iT Broad Range RNA Assay kit (Life Technologies), respectively.
Vornholt E., Drake J., Mamdani M., McMichael G., Taylor Z.N., Bacanu S., Miles M.F, & Vladimirov V.I. (2021). Identifying a novel biological mechanism for alcohol addiction associated with circRNA networks acting as potential miRNA sponges. Addiction Biology, 26(6), e13071.
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6

Quantitative Analysis of LOC339975 in AD

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PFC and NAc tissues from 41 AD cases and 41 controls were provided by
the New South Wales Tissue Resource Centre. Age, sex,
ethnicity, brain weight, brain pH, post-mortem interval (PMI), tissue
hemisphere, cause of death, blood toxicology, smoking status, neuropathology and
liver pathology were provided for each subject. Confounding effects of all these
covariates were controlled by analysis of covariance (ANCOVA, Supplementary Table S7).
Total RNA was isolated from 100mg frozen tissue using the mirVana-PARIS kit
(Life Technologies, Carlsbad, CA) following manufacturer’s protocols.
RNA concentration was measured using the Quant-iT Broad Range RNA Assay kit
(Life Technologies). The RNA Integrity Number (RIN) was measured on the Agilent
2100 Bioanalyzer (Agilent Technologies, Inc., Santa Clara, CA). Quantitative
real-time PCR (qRT-PCR) analyses were performed as previously described using
SYBR Green (Riley et al., 2010 (link)) with
primers spanning the LOC339975 exon 2–3 junction.
Samples with missing genotypes and outliers (±2SD from the mean) were
omitted from further analysis.
Adkins A.E., Hack L.M., Bigdeli T.B., Williamson V.S., McMichael G.O., Mamdani M., Edwards A., Aliev F., Chan R.F., Bhandari P., Raabe R.C., Alaimo J.T., Blackwell G.G., Moscati A.A., Poland R.S., Rood B., Patterson D.G., Walsh D., Whitfield J.B., Zhu G., Montgomery G.W., Henders A.K., Martin N.G., Heath A.C., Madden P.A., Frank J., Ridinger M., Wodarz N., Soyka M., Zill P., Ising M., Nöthen M.M., Kiefer F., Rietschel M., Gelernter J., Sherva R., Koesterer R., Almasy L., Zhao H., Kranzler H.R., Farrer L.A., Maher B.S., Prescott C.A., Dick D.M., Bacanu S.A., Mathies L.D., Davies A.G., Vladimirov V.I., Grotewiel M., Bowers M.S., Bettinger J.C., Webb B.T., Miles M.F., Kendler K.S, & Riley B.P. (2017). Genomewide Association Study of Alcohol Dependence Identifies Risk Loci Altering Ethanol-response Behaviors in Model Organisms. Alcoholism, clinical and experimental research, 41(5), 911-928.
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7

Extraction and Quantification of Total RNA

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Total RNA was isolated from 50mg of frozen tissue from NAc using the mirVana-PARIS kit (Thermo Fisher, Carlsbad, CA), following manufacturer’s protocols. RNA concentration was measured using the Quant-iT Broad Range RNA Assay kit (Life Technologies), and the RNA Integrity Number (RIN) was measured on the Agilent 2100 Bioanalyzer (Agilent Technologies, Inc., Santa Clara, CA).
Drake J., McMichael G.O., Vornholt E.S., Cresswell K., Williamson V., Chatzinakos C., Mamdani M., Hariharan S., Kendler K.S., Kalsi G., Riley B.P., Dozmorov M., Miles M.F., Bacanu S, & Vladimirov V.I. (2020). Assessing the Role of Long Noncoding RNA in Nucleus Accumbens in Subjects With Alcohol Dependence. Alcoholism, Clinical and Experimental Research, 44(12), 2468-2480.
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8

Total RNA Isolation and cDNA Synthesis

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Total RNA from the frozen tissue was isolated using acidic phenol chloroform extraction and alcohol precipitation method (18 (link)). RNA quantity was measured by Quant-iT™ RNA broad range assay kit (Invitrogen, Carlsbad, USA) and Qubit® 2.0 Fluorometer (Life Technologies, Carlsbad, USA), and its integrity was confirmed by 1% agarose gel electrophoresis. Reverse transcription was carried out using QuantiTect reverse transcription kit (Qiagen GmbH, Hilden, Germany) with 1000 ng of total RNA in a 20 μl reaction volume, as mentioned in the manufacturer’s protocol. The cDNA obtained were subjected to 10-fold dilution, before being used in qPCR.
Vasanth G., Kiron V., Kulkarni A., Dahle D., Lokesh J, & Kitani Y. (2015). A Microbial Feed Additive Abates Intestinal Inflammation in Atlantic Salmon. Frontiers in Immunology, 6, 409.
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9

Mouse Myeloid Innate Immunity Gene Expression

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Spleens were incubated in RNAlater (Thermo Fisher Scientific), followed by homogenization in a Geno/Grinder (SPEX SamplePrep, Metuchen, NJ, USA) using ZR BashingBead Lysis Tubes (Zymo Research, Irvine, CA, USA). RNA was isolated using automated Maxwell RSC Instrument with simplyRNA Tissue Kit as per the manufacturer’s recommendations (Promega Corp., Madison, WI, USA). RNA quality was assessed using TapeStation RNA ScreenTape (Agilent, Santa Clara, CA, USA) and quantitated using the Quant-iT RNA Broad Range Assay Kit (Invitrogen). RNA (200 ng) was assessed for gene expression using the nCounter Mouse Myeloid Innate Immunity v2 Panel (NanoString Technologies, Seattle, WA, USA) according to the nCounter XT CodeSet Gene Expression assay protocol. Resulting RCC files were examined with NanoString nSolver software for quality control analysis.
Nelson J., Sorensen E.W., Mintri S., Rabideau A.E., Zheng W., Besin G., Khatwani N., Su S.V., Miracco E.J., Issa W.J., Hoge S., Stanton M.G, & Joyal J.L. (2020). Impact of mRNA chemistry and manufacturing process on innate immune activation. Science Advances, 6(26), eaaz6893.
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10

Automated RNA Isolation and NanoString Analysis

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RNA was isolated from spinal cord tissue using automated Promega Maxwells with the RSC simplyRNA Tissue Kit as per the manufacturer’s recommendations (Promega, cat #AS1340). RNA quality was assessed using Agilent Tape Station RNA ScreenTape (Agilent, cat #5067-5576) and quantitated by Invitrogen Quant-iT RNA Broad Range Assay Kit (Invitrogen, cat #Q10213). 150 ng of RNA was used as input for NanoString Autoimmune Panel (NanoString cat #XT-CSO-MAIP1-12) according to nCounter XT CodeSet Gene Expression Assays protocol. Resulting RCC files were examined with NanoString nSolver software for QC analysis. Briefly, samples had a background threshold set to 25 counts and normalized to a set of housekeeping genes with low variance (%CV < 40%) and medium or high counts (>50).
de Picciotto S., DeVita N., Hsiao C.J., Honan C., Tse S.W., Nguyen M., Ferrari J.D., Zheng W., Wipke B.T, & Huang E. (2022). Selective activation and expansion of regulatory T cells using lipid encapsulated mRNA encoding a long-acting IL-2 mutein. Nature Communications, 13, 3866.
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