Dna eraser

Manufactured by Takara Bio

Sourced in Japan

The DNA Eraser is a laboratory instrument designed to remove DNA from various surfaces and equipment. It utilizes a specialized cleaning process to ensure effective decontamination and prevent cross-contamination in research and diagnostic settings.

Automatically generated - may contain errors

Lab products found in correlation

Primescript rt reagent kit, by Takara Bio (6 mentions) All in one mirna qrt pcr detection kit, by GeneCopoeia (4 mentions) Trizol reagent, by Thermo Fisher Scientific (4 mentions) Ribozol, by Avantor (2 mentions) Quantitect sybr green pcr kit, by Qiagen (2 mentions) Applied biosystems 7500 real time pcr system, by Thermo Fisher Scientific (2 mentions) Nanodrop 1000 spectrophotometer, by Thermo Fisher Scientific (2 mentions) Quantstudio 3 real time pcr system, by Thermo Fisher Scientific (1 mentions) Powertrack green master mix, by Thermo Fisher Scientific (1 mentions) Spss v17, by IBM (1 mentions) Primer premier 5, by Premier Biosoft (1 mentions) Reverse transcription kit, by Thermo Fisher Scientific (1 mentions) Revertra ace qpcr rt kit, by Toyobo (1 mentions) Rnazol reagent, by Merck Group (1 mentions) Sybr green quantitative rt qpcr kit, by Merck Group (1 mentions) Cfx96 touch real time pcr detection system, by Bio-Rad (1 mentions) Primescripttm rt reagent kit, by Takara Bio (1 mentions) Rnase free dnase 1, by Takara Bio (1 mentions) Nd 1000 spectrophotometer, by Thermo Fisher Scientific (1 mentions) Primescript rt polymerase, by Takara Bio (1 mentions)

15 protocols using dna eraser

Reverse Transcription and qPCR Analysis

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Using the PrimeScript™ RT reagent kit (CAS No.: RR037B, Takara), reverse transcription was performed to transform RNAs into cDNA with high quality and at high quantity. Subsequently, DNA Eraser (Takara) was used to eliminate the genomic DNA in RNA, and then β-actin primers were used as a house gene to evaluate cDNA quality.
The specific DEG primers were designed by Primer Premier 5.0 (Premier Biosoft International, USA; Supplementary Table S1). The cDNA product was subjected to qPCR on the QuantStudio™ 3 Real-Time PCR System (CAS No.: A28136, ABI, United States). Each reaction system included 1 μL cDNA template, 0.5 μL forward primer, 0.5 μL reverse primer, 5 μL fluorescent dye (PowerTrack Green Master Mix, CAS No.: A46112, ABI, United States), and 3 μL ddH₂O. The reaction was performed under the following conditions: 50°C for 2 min, 95°C for 10 min, followed by 40 amplification cycles (95°C for 15 s and 60°C for 45 s). After amplification, the specificity of PCR was detected using the melting curve, and relative expression was calculated by 2^−ΔΔCt method (18 (link)). Finally, t-test was performed to test for significant differences between genes in the CL and GL groups in SPSS v17.0 (SPSS, Chicago, IL, United States).

Zhong H., Lou C., Ren B., Pi J., Dai T., Qin W, & Zhou Y. (2023). Hepatic transcriptome analysis provides new insights into ghrelin regulation of the liver in Nile tilapia (Oreochromis niloticus). Frontiers in Veterinary Science, 10, 1192195.

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Profiling Circulating RNA Integrity

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Total RNAs in plasma and HBEpCs were extracted using Ribozol (VWR) and digested with a DNA eraser (Takara, Japan) until all samples reached an OD260/280 ratio close to 2.0, which indicated pure RNA. Electrophoresis (5% urea-PAGE gel) was carried out to analyze the integrity of RNA samples. Only RNA samples with high purity and satisfactory integrity were subjected to subsequent assays. The cDNA was synthesized using reverse transcription kit (Fermentas, USA). RT-qPCR assay was performed using ReverTra Ace™ qPCR RT Kit (Toyobo, Japan). GAPDH and U6 were selected as internal controls. Ct values of circFADS2 and miR-133a were normalized to their endogenous controls using the 2^-ΔΔCt method. The primer sequences were presented in Supplementary Table 1.

Niu F., Liang X., Ni J., Xia Z., Jiang L., Wang H., Liu H., Shen G, & Li X. (2022). CircRNA circFADS2 is under-expressed in sepsis and protects lung cells from LPS-induced apoptosis by downregulating miR-133a. Journal of Inflammation (London, England), 19, 4.

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Quantitative Analysis of Circular RNA and MicroRNA

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Total RNA was isolated from BMMNCs and in vitro cultured cells using RNAzol reagent (Sigma-Aldrich; Merck KGaA), and treated with DNA eraser (Takara Bio, Inc.) at 37°C until the OD260 nm/280 nm ratio had reached ~2.0 (pure RNA). The integrity of all RNA samples was determined by electrophoresis using a 5% urea-PAGE gel. Total RNA was used for first-strand cDNA synthesis with the Prime Script RT reagent kit (Takara Bio, Inc.) per the manufacturer's instructions. Circ-ATAD1 expression was determined using the SYBR^® Green Quantitative RT-qPCR Kit (Sigma-Aldrich; Merck KGaA) with GAPDH as the internal control. Mature miR-34b expression was analyzed using the All-in-One™ miRNA qRT-PCR Detection Kit (GeneCopoeia, Inc.) with U6 as the internal control. All operations were completed following the manufacturers' instructions. The qPCR thermocycling conditions for all genes, cirRNA and miRNAs were as follows: 95°C for 1 min, followed by 40 cycles of 95°C for 10 sec and 60°C for 45 sec. PCR reactions were performed using the CFX96 Touch Real-Time PCR Detection System (Bio-Rad Laboratories, Inc.). Ct values of the targeted genes were normalized to the corresponding internal controls based on the 2^−ΔΔCq method, which was used to quantify gene expression (18 (link)). The qPCR primers are listed in Table II.

Wu Y., Gao B., Qi X., Bai L., Li B., Bao H., Wu X., Wu X, & Zhao Y. (2021). Circular RNA ATAD1 is upregulated in acute myeloid leukemia and promotes cancer cell proliferation by downregulating miR-34b via promoter methylation. Oncology Letters, 22(5), 799.

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Endometrial RNA Extraction and Analysis

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The Trizol reagent (Invitrogen, Carlsbad, CA, USA) was used to extract the RNA from the endometrium of each cow. The RNA concentration was evaluated with an ND-1000 UV-vis spectrophotometer (NanoDrop Ltd., Wilmington, DE, USA), and the quality of RNA was determined by both 1% agarose gel electrophoresis and the 260/280 absorbanceratio using an ND1000 spectrophotometer (NanoDrop Technologies Inc., Wilmington, DE, USA) [14 (link)]. The subsequent experiments only used the RNA samples with a ratio from 1.8–2.1. Next, 5× gDNA Eraser Buffer, 2.0 μL Eraser Buffer, 1.0 μL DNA Eraser and RNase-free DNase I (TaKaRa, Dalian, China) were used to remove the chromosomal DNA in 1 µg of RNA samples (42 °C, 2 min). Thereafter, a reverse transcription was conducted using a PrimeScript^TM RT reagent kit (TaKaRa, Japan), and stored at −80 °C for real-time polymerase chain reaction (RT-PCR) analysis.

Wang Y., Han X., Tan Z., Kang J, & Wang Z. (2020). Rumen-Protected Glucose Stimulates the Insulin-Like Growth Factor System and mTOR/AKT Pathway in the Endometrium of Early Postpartum Dairy Cows. Animals : an Open Access Journal from MDPI, 10(2), 357.

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LUADT1, Pim-1, and miR-195 Expression Analysis

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Total RNAs were extracted from both tissue samples and in vitro cultured cells using Trizol reagent (Invitrogen). For LPS treatment, HCAECs were cultured in cell culture medium containing 0, 100,150, 200 and 500 ng/ml LPS for 24 h before use. DNA eraser (Takara) was used to digest all RNA samples to remove genomic DNA. Reverse transcriptions (RTs) were performed using PrimeScript RT-polymerase (Takara) with total RNA samples as template. With cDNA samples as template, qPCR reaction mixtures were prepared using QuantiTect SYBR Green PCR Kit (QIAGEN). The expression levels of LUADT1 and Pim-1 mRNA were measured with GAPDH as endogenous control. Extraction of miRNAs from aforementioned samples and cells was performed using miRNeasy Mini Kit (QIAGEN). The expression levels of mature miR-195 were measured using All-in-One™ miRNA qRT-PCR Detection Kit (Genecopoeia) with U6 as endogenous control. All PCR reactions were performed in triplicate manner and fold changes of gene expression levels were calculated using 2^−ΔΔCt method.

Zhang Z., Lv M., Wang X., Zhao Z., Jiang D, & Wang L. (2020). LncRNA LUADT1 sponges miR-195 to prevent cardiac endothelial cell apoptosis in sepsis. Molecular Medicine, 26, 112.

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Quantifying TONSL-AS1, CDK1, and miR-490-3p

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TRIZOL reagent (Invitrogen) was used to extract total RNAs. With 85% ethanol used in RNA precipitation and washing, miRNAs were retained. Genomic DNAs were removed by incubating RNA samples with DNA eraser (Takara, Japan) at 37 °C for 2 h. First strand cDNA synthesis kit (Takara) was used for reverse transcriptions (RTs) with poly (T) as primer and RNA samples as template. With cDNA at template, qPCR mixtures were prepared using SYBR Green dye (Takara). Expression levels of TONSL-AS1 and CDK1 mRNA were normalized with GAPDH as endogenous control. Measurement of the expression levels of miR-490-3p was performed using mirVana qRT-PCR miRNA Detection Kit (Thermo Fisher Scientific) with all operations performed following the manufacturer’s instructions. Fold changes of gene expression were normalized using 2^-ΔΔCt method.

Liu Y., Li L., Wang X., Wang P, & Wang Z. (2020). LncRNA TONSL-AS1 regulates miR-490-3p/CDK1 to affect ovarian epithelial carcinoma cell proliferation. Journal of Ovarian Research, 13, 60.

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Quantification of lncRNA Expression

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Briefly, cDNA was transcribed from total RNA using the PrimeScript RT reagent kit with a DNA Eraser (TaKaRa, Shiga, Japan). Primers for lncRNAs were designed and synthesized. Then, qPCR assays were performed using a Bio-Rad IQTM5 Multicolor Real-Time qRT-PCR detection system (Bio-Rad, Hercules, CA, USA). The expression levels of lncRNAs and genes were detected using primers specific for the lncRNAs and mRNAs (Additional file 1: Table S1). Human beta-actin was used as a housekeeping gene for normalization. The expression levels of lncRNAs and mRNAs were measured in terms of the cycle threshold (CT) and normalized to beta-actin expression using the 2^-ΔΔCt method.

Li G., Liu Y., Liu C., Su Z., Ren S., Wang Y., Deng T., Huang D., Tian Y, & Qiu Y. (2016). Genome-wide analyses of long noncoding RNA expression profiles correlated with radioresistance in nasopharyngeal carcinoma via next-generation deep sequencing. BMC Cancer, 16(1), 719.

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Gene Expression Analysis of Tibial Bone

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Bilateral tibias were homogenized in liquid nitrogen to collect the mRNA samples. Total RNA was extracted using Trizol (Invitrogen, Carlsbad, USA) following the manufacturer's protocol. Then, cDNA was synthesized using the Prime Script RT reagent kit with DNA Eraser (Takara Bio, Shiga, Japan). Equal volumes of cDNA were amplified in a real-time detection system (ABI7500, USA) with FastStart Universal SYBR Premix (Yeasen, Shanghai, China) and primers (Sangon Biotech, Shanghai, China). Relative mRNA expression was calculated using the relative standard curve method (2^−ΔΔCT) with β-actin as the endogenous control. The primer sequence of RANK, ALP, collagen type I (Col-1), runt-related transcription factor 2 (Runx2), osteocalcin (OCN), osteoprotegerin (OPG), RANKL, NFATC1, and c-fos are listed in online supplementary Table 1 (for all online suppl. material, see www.karger.com/doi/10.1159/000527502).

Huang Y.M., Xu B., Kuai Z., He Y.T., Lu Y., Shen J.P., Wu K.F., Wu J.Y., Ren W.Y, & Hu Y. (2022). Glucagon-Like Peptide-2 Ameliorates Age-Associated Bone Loss and Gut Barrier Dysfunction in Senescence-Accelerated Mouse Prone 6 Mice. Gerontology, 69(4), 428-449.

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Quantification of miRNA and mRNA Expression

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Ribozol (Sigma-Aldrich) was used to perform all RNA extractions following the manufacture’s instructions. RNA samples were precipitated using 85% ethanol to harvest miRNAs. All RNA samples were digested by DNA eraser (Takara, USA). SSRT IV system (Thermo Fisher Scientific) was used to transcribe total RNAs into cDNAs. With cDNA samples as template, SYBR® Green Realtime PCR Master Mix (Toyobo, Japan) was used to prepare all qPCR mixtures. GAPDH was used as the endogenous control to measure the expession levels of MIR17HG and Bach-1. All-in-One™ miRNA qRT-PCR Detection Kit (GeneCopoeia) was used to measure the expression levels of mature miR-142-3p. All steps including polyadenylation, reverse transcriptions and qPCR assays were performed following the instructions from GeneCopoeia. U6 was used as the endogenous control. All qPCR assays were repeated 3 times and data normalizations were performed using 2^-ΔΔCq method.

Wei S., Liu J., Li X, & Liu X. (2020). LncRNA MIR17HG inhibits non-small cell lung cancer by upregulating miR-142-3p to downregulate Bach-1. BMC Pulmonary Medicine, 20, 78.

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Cardiac Biomarker Quantification in Rats

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Specimens from left ventricular tissues of rats in each group were weighed and cut into pieces, mixed with pre-cooled PBS at a ratio of 1:9 (weight:volume) to prepare the tissue homogenate. Myocardial levels of renin (Sigma-Aldrich; Merck KGaA; cat. no. RAB1162), TNF-α (Sigma-Aldrich; Merck KGaA; cat. no. RAB0479), IL-6 (Sigma-Aldrich; Merck KGaA; cat. no. RAB0311), brain natriuretic peptide (BNP; Sigma-Aldrich; Merck KGaA; cat. no. RAB0386), aldosterone (ALD; Abcam; cat. no. Ab136933), malondialdehyde (MDA; Abcam; cat. no. Ab238537), ROS (Jianglai Bio; http://www.laibio.com/; cat. no. JL21051), superoxide dismutase (SOD; Jianglai Bio; cat. no. JL11065), NOX1 (Jianglai Bio; cat. no. JL36449), NOX2 (Jianglai Bio; cat. no. JL50110) and NOX4 (Jianglai Bio; cat. no. JL23194) were measured. ELISA was performed according to the manufacturer's protocol and a DNA Eraser was used (Takara Bio, Inc.).

Cheng D., Chen L., Tu W., Wang H., Wang Q., Meng L., Li Z, & Yu Q. (2020). Protective effects of valsartan administration on doxorubicin-induced myocardial injury in rats and the role of oxidative stress and NOX2/NOX4 signaling. Molecular Medicine Reports, 22(5), 4151-4162.

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