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Dnase reagents

Manufactured by Thermo Fisher Scientific
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DNase reagents are laboratory products used to degrade DNA molecules. They are designed to remove DNA from biological samples, facilitating downstream applications that require DNA-free environments.

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Lab products found in correlation

Rnaaqueous, by Thermo Fisher Scientific (2 mentions) Rnalater stabilizing solution, by Thermo Fisher Scientific (1 mentions) Rnaqueous 4pcr total rna isolation kit, by Thermo Fisher Scientific (1 mentions) Lysing matrix d, by MP Biomedicals (1 mentions) Fastprep 24, by MP Biomedicals (1 mentions) Truseq mrna library kit, by Illumina (1 mentions) Agilent 2100 bioanalyzer, by Agilent Technologies (1 mentions)

3 protocols using dnase reagents

1

Porcine Salivary Gland Gene Expression

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Tissue samples for real-time PCR analysis were collected from three separate regions of the porcine PG and stored in RNAlater stabilizing solution (Ambion, Houston, TX, USA). Tissue (∼25 mg) was homogenized in RNA lysis buffer using Fastprep-24 instrument with lysing matrix D (MP Biomedicals, Irvine, CA, USA). Murine SMGs were harvested, minced, and lyzed by homogenization. RNA was subsequently isolated using the RNAqueous-4PCR total RNA isolation kit and DNase reagents according to the manufacturer’s instructions (Ambion, Houston, TX, USA). cDNA was generated from DNase-free RNA samples, amplified, and gene expression was normalized to the housekeeping gene RPS29 or Gapdh as previously described in Patel et al.40 (link)
Lombaert I.M., Patel V.N., Jones C.E., Villier D.C., Canada A.E., Moore M.R., Berenstein E., Zheng C., Goldsmith C.M., Chorini J.A., Martin D., Zourelias L., Trombetta M.G., Edwards P.C., Meyer K., Ando D., Passineau M.J, & Hoffman M.P. (2020). CERE-120 Prevents Irradiation-Induced Hypofunction and Restores Immune Homeostasis in Porcine Salivary Glands. Molecular Therapy. Methods & Clinical Development, 18, 839-855.
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2

RNA Extraction and Sequencing

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Total RNA was collected and purified using RNAaqueous and DNase reagents according to the manufacturer’s instructions (Ambion, Houston, TX, USA). RNA quality was assessed using the Agilent 2100 BioAnalyzer, and samples with an RNA integrity number ≥ 6.0 were included for RNA sequencing. Up to 1 μg RNA was used to synthesize mRNA libraries using the TruSeq mRNA library kit (Illumina Inc, San Diego, CA, USA), as per the manufacturer’s instructions.
Efraim Y., Chen F.Y., Stashko C., Cheong K.N., Gaylord E., McNamara N, & Knox S.M. (2020). Alterations in corneal biomechanics underlie early stages of autoimmune-mediated dry eye disease. Journal of autoimmunity, 114, 102500.
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3

Quantitative Real-Time PCR Analysis

Cited 1 time
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Real-time PCR was performed as previously described [26 (link)]. Total RNA was collected and purified using RNAaqueous and DNase reagents according to manufacture’s protocol (Ambion, Houston, TX). cDNA prepared using Superscript reagents (BioRad, CA) and SYBR-green qPCR was performed using 5ng of cDNA, with primers designed using Beacon Designer software. Target gene expression was normalized using eukaryotic 18s rRNA as an endogenous control. All reactions were performed in triplicate and repeated three times.
Chen F.Y., Lee A., Ge S., Nathan S., Knox S.M, & McNamara N.A. (2017). Aire-deficient mice provide a model of corneal and lacrimal gland neuropathy in Sjögren's syndrome. PLoS ONE, 12(9), e0184916.
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