Deoxyribonuclease 1 treatment

Manufactured by Qiagen

Deoxyribonuclease I (DNase I) is an enzyme used to degrade DNA. It catalyzes the hydrolytic cleavage of phosphodiester linkages in the DNA backbone, resulting in the formation of 5'-phosphate and 3'-hydroxyl termini. This process is used in various molecular biology applications to remove unwanted DNA from samples.

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Lab products found in correlation

Taqman probe, by Thermo Fisher Scientific (4 mentions) Taqman gene expression master mix 2 with ung, by Thermo Fisher Scientific (2 mentions) Rneasy micro kit, by Qiagen (2 mentions) Nanodrop 1000, by Thermo Fisher Scientific (2 mentions) High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (2 mentions) 7900ht fast real time pcr system, by Thermo Fisher Scientific (1 mentions) Iscript cdna synthesis kit, by Bio-Rad (1 mentions) Mikro dismembrator u, by Sartorius (1 mentions) Phase lock gel, by Quantabio (1 mentions) Quantstudio 6 flex system, by Thermo Fisher Scientific (1 mentions) Rneasy mini kit, by Qiagen (1 mentions) Cdna synthesis kit, by Promega (1 mentions)

Close spelling variants of this product

Deoxyribonuclease treatment Deoxyribonuclease I Deoxyribonuclease

3 protocols using deoxyribonuclease 1 treatment

Quantifying Gene Expression with Real-Time PCR

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RNA was isolated using Trizol and the RNeasy micro kit with on-column deoxyribonuclease I treatment (Qiagen). Quality and integrity of RNA was determined using nanodrop spectrophotometer ND-1000. Reverse-transcriptase reactions were prepared using 1 µg of RNA and iScript cDNA Synthesis Kit (Bio-Rad). Quantitative real-time PCR was completed using TaqMan probes (Applied Biosystems) for Gcg (Mm01269055_m1), Agrp (Mm00475829_g1), Kiss1 (Mm03058560_m1), and housekeeping gene 18s (Hs03003631_g1) was used as an endogenous control to normalize each sample and gene. PCRs were in a 10-μl volume using 0.5 μl TaqMan probe, 10 ng cDNA template, 5 μl TaqMan Gene Expression Master Mix II with UNG (Applied Biosystems), and 2.5 μl DNase/RNase-free molecular grade water (Qiagen). Real-time PCR was run using an Applied Biosystems 7900HT Fast Real-Time PCR system with initial denaturing at 50°C for 2 min and 95°C for 10 min, followed by 40 cycles at 95°C for 15 s, and annealing at 60°C for 1 min. Results were calculated using the Pfaffl method (Pfaffl, 2001 (link)).

Heppner K.M., Baquero A.F., Bennett C.M., Lindsley S.R., Kirigiti M.A., Bennett B., Bosch M.A., Mercer A.J., Rønnekleiv O.K., True C., Grove K.L, & Smith M.S. (2017). GLP-1R Signaling Directly Activates Arcuate Nucleus Kisspeptin Action in Brain Slices but Does not Rescue Luteinizing Hormone Inhibition in Ovariectomized Mice During Negative Energy Balance. eNeuro, 4(1), ENEURO.0198-16.2016.

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Murine Cartilage Gene Expression Analysis

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Humeral head articular cartilage gene expression was determined in 8 BtxA (4M, 4F), 8 Saline (4F, 4M), and 8 Control (4M, 4F) shoulders from 40-week old mice using established methods.³² Immediately following mouse sacrifice and dissection, tissues were frozen in liquid nitrogen and pulverized using a ball mill homogenizer (Mikro-Dismembrator U, Sartorius). RNA extraction was performed using guandinium thiocyanatephenol-chloroform (TRIzol, Thermo Fisher Scientific) and interphase separation (Phase Lock Gel, QuantaBio). RNA cleanup was performed using spin columns (RNeasy Mini Kit, Qiagen) with on-column deoxyribonuclease I treatment (Qiagen). RNA quantity and quality were determined using a spectrophotometer (NanoDrop 1000, Thermo Fisher Scientific). RNA was then reverse-transcribed into cDNA using a High-Capacity cDNA Reverse Transcription Kit (Thermo Fisher Scientific). Cartilage-, bone- and OA-related genes (Ihh, Acan, Runx2, Dkk1, Col2A1, Col10A1, BGLAP, ALPL, and BMP2) were evaluated by SYBR Green-based quantitative reverse transcription-polymerase chain reaction on an Applied Biosystems Quant-Studio 6 flex system. Glycer-aldehyde 3-phosphate dehydrogenase served as a housekeeping gene, and the relative mRNA expression level for each target gene was determined as 2^-ΔΔCt.

Forrester L.A., Fang F., Jacobsen T., Hu Y., Kurtaliaj I., Roye B.D., Guo X.E., Chahine N.O, & Thomopoulos S. (2021). Transient neonatal shoulder paralysis causes early osteoarthritis in a mouse model. Journal of orthopaedic research : official publication of the Orthopaedic Research Society, 40(9), 1981-1992.

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Quantifying Gene Expression in Murine Hypothalamus

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Hypothalami were collected from control and DIO mice. Tissue was homogenized in Trizol and RNA was isolated using the RNeasy micro kit with on-column deoxyribonuclease I treatment (Qiagen). Quality and integrity of RNA was determined using a ND-1000 Nanodrop spectrophotometer. Reverse-transcription reactions were prepared using 2 μg of RNA and a cDNA Synthesis Kit (Promega). Quantitative real-time PCR was performed using TaqMan probes (Applied Biosystems) for RELN (Mm00465200_m1), ApoER2 (Mm00474030_m1), and VLDLR (Mm00443298_m1). The level of 18s rRNA (Hs03003631_g1) was used as an endogenous control for normalization. PCRs were performed in a 10-μl volume using 0.5 μl TaqMan probe, 20 ng cDNA template, 5 μl TaqMan Gene Expression Master Mix II with UNG (Applied Biosystems), and 2.5 μl DNase/RNase molecular-grade water (Qiagen). Real-time PCR was run using a 7900HT Fast Real-Time PCR system with initial denaturation at 50 °C for 2 min and 95 °C for 10 min, followed by 40 cycles at 95 °C for 15 s, and annealing at 60 °C for 1 min.

Roberts B.L., Bennett B.J., Bennett C.M., Carroll J.M., Dalbøge L.S., Hall C., Hassouneh W., Heppner K.M., Kirigiti M.A., Lindsley S.R., Tennant K.G., True C.A., Whittle A., Wolf A.C., Roberts CT J.r., Tang-Christensen M., Sleeman M.W., Cowley M.A., Grove K.L, & Kievit P. (2019). Reelin is modulated by diet-induced obesity and has direct actions on arcuate proopiomelanocortin neurons. Molecular Metabolism, 26, 18-29.

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