Agarose, PIPES, Potassium hydroxide, Tween® 20, 2-mercaptoethanol, Trypsin, MPH, ATX and VPA were purchased from Sigma (St. Louis, MO, USA). ECLTM Western blotting detection reagents were obtained from Amersham Life Science (Arlington Heights, IL, USA). Trizol and SuperScriptTM II Reverse Transcriptase were purchased from Invitrogen (Carlsbad, CA, USA). DNase Ι was purchased from Roche (Mannheim, Germany). Protein G Agarose was obtained from Millipore Corporation (Billerica, MA, USA). Taq polymerase and dNTP were obtained from Takara (Shiga, Japan). Protease inhibitor cocktail was obtained from Calbiochem (La Jolla, CA, USA). Chelex 100 was obtained from BioRad (Hercules, CA, USA). Antibodies were purchased from the following companies: anti-β-actin from Sigma (St. Louis, MO, USA), Histone H3, Acetyl-Histone H3 and HDAC1 antibody from Cell signaling (Boston, MA, USA), DAT, NET and peroxidase-conjugated secondary antibody from Santa Cruz biotechnology (Dallas, TX, USA).
Dnase ι
Manufactured by Roche
Sourced in Germany
DNase I is an enzyme that catalyzes the hydrolytic cleavage of DNA. It is a single-stranded DNA-specific endonuclease that randomly cleaves the phosphodiester bonds within the DNA backbone, resulting in the generation of oligonucleotides with 5'-phosphate and 3'-hydroxyl termini.
Automatically generated - may contain errors
Lab products found in correlation
Agarose, by Merck Group (1 mentions)
Taq polymerase, by Takara Bio (1 mentions)
Protein g agarose, by Merck Group (1 mentions)
Peroxidase conjugated secondary antibody, by Santa Cruz Biotechnology (1 mentions)
Ecltm western blotting detection reagent, by Cytiva (1 mentions)
Hdac1 antibody, by Cell Signaling Technology (1 mentions)
Histone h3, by Cell Signaling Technology (1 mentions)
Pipes, by Merck Group (1 mentions)
Superscript 2 reverse transcriptase, by Thermo Fisher Scientific (1 mentions)
Acetyl histone h3, by Cell Signaling Technology (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
Tween 20, by Merck Group (1 mentions)
Trypsin, by Merck Group (1 mentions)
Anti β actin, by Merck Group (1 mentions)
Chelex 100, by Bio-Rad (1 mentions)
Rnase free water, by Thermo Fisher Scientific (1 mentions)
High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (1 mentions)
2720 thermal cycler, by Thermo Fisher Scientific (1 mentions)
Tri reagent, by Merck Group (1 mentions)
2 protocols using dnase ι
1
Molecular Investigations in Neuroscience
2
RNA Extraction and Reverse Transcription
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Frozen liver samples (50–100 mg) were ground using mortar and pestle with liquid nitrogen, and total RNA was extracted with TRI reagent (Sigma, St. Louis, MO). RNA samples were incubated with DNase Ι (Roche, Mannheim, Germany) at room temperature for 20 min, followed by 10 min in 90 °C heat block to prevent contamination of genomic DNA. High-capacity cDNA Reverse Transcription Kit (Applied Biosystems, Foster City, CA) was used for reverse transcription of 2 μg of total RNA. Reverse transcription was performed in a 2720 Thermal Cycler (Applied Biosystems) with 20 μL reaction volume with the following program: heated at 25 °C for 10 min, incubated at 37 °C for 2 h, and then heated at 85 °C for 5 s. The 20 μL of final product was diluted to 400 μL for 5 ng/μL using RNase free water (Fisher, Fair Lawn, NJ) and stored at − 20 °C.
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