K562 cells were incubated with different doses of molecule L20 and SAHA for 24 h. After treatment, cells were collected and fixed with 70% pre-cold ethanol in PBS and stored at −20 °C overnight. Then washed the cells with PBS twice, and incubated with 100 µg/mL RNase I (Solarbio, China) at 37 °C for 1 h, stained with propidium iodide (PI, 10 µg/mL, Solarbio, China) for 30 min avoid light at room temperature. Finally, DNA content was measured by flow cytometry (FACSAriaIII, Becton Dickinson, USA). The data were analysed and fitted by ModFit software.
Rnase 1
Manufactured by Solarbio
Sourced in China
RNase I is a ribonuclease enzyme that catalyzes the hydrolysis of single-stranded RNA. It is used in molecular biology for the removal of unwanted RNA from DNA samples.
Automatically generated - may contain errors
5 protocols using rnase 1
1
Cell Cycle Analysis of K562 Cells
2
Vesicular Nucleic Acid and Protein Removal
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To remove vesicular nucleic acids and proteins, 1 mg/mL of native LrEVs were treated with 10 U/mL of DNase I (Thermo Scientific, #EN0521), 10 U/mL of RNase I (Solarbio, #R8021), and 1 mg/mL of proteinase K-agarose (Sigma, #P9290), respectively, as described previously [16 (link), 31 (link)]. The native LrEVs were incubated with DNase I and RNase I at 37 °C for 30 min, and with proteinase K-agarose at 37 °C for 2 h. The enzymes were inactivated at 75 °C for 1 h. proteinase K-agarose was additionally removed by centrifugation (12,000×g, 1 min).
3
Cell Cycle Analysis of Molecule C2 Effects
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K562 and HCC1937 cells were incubated with different doses of molecule C2 , olaparib, or chlorambucil for 24 h. After treatment, cells were collected and fixed with 70% pre-chilled ethanol in PBS and stored at −20°C overnight. The cells were then washed twice with PBS, incubated with 100 μg/ml RNase I (Solarbio, China) at 37°C for 1 h, and stained with propidium iodide (PI, 10 μg/ml, Solarbio, China) for 30 min, in the dark, at room temperature. Finally, the DNA content was measured using flow cytometry (FACSAria III, Becton Dickinson, United States). The data were analyzed and fitted using ModFit software.
4
Cell Cycle Analysis of K562 Cells
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K562 cells were incubated with different doses of molecule D28 and SAHA for 24 h. After treatment, cells were collected and fixed with 70% pre-cold ethanol in PBS and stored at −20°C overnight. Then, the cells were washed with PBS twice and incubated with 100 μg/ml RNase I (Solarbio, China) at 37°C for 1 h and stained with propidium iodide (PI, 10 μg/ml, Solarbio, China) for 30 min avoiding light at room temperature. Finally, DNA content was measured by flow cytometry (FACSAriaIII, Becton Dickinson, United States). The data was analyzed and fitted by ModFit software.
5
Phage Genome Extraction and Annotation
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The genome was extracted from the phage with the Tris-HCL method (20 (link)). The genome was digested with DNase I, RNase I, and mung bean nuclease (Solarbio) to determine the genome type. The phage genome was scanned and sequenced with Illumina sequencing technology. The databases RefSeq nonredundant (Nr) protein (RefSeq: NCBI Reference Sequence Database, nih.gov), Clusters of Orthologous Groups of proteins (COG, https://www.ncbi.nlm.nih.gov/research/cog ), Kyoto Encyclopedia of Genes and Genomes (KEGG, https://www.kegg.jp/kegg/ ), Swiss-Prot (www.sib.swiss/swiss -prot), and Gene Ontology (http://www.geneontology.org/GO.indices.html ) were used to annotate the genes. Finally, the SnapGene software (Insightful Science; available at snapgene.com) was used to analyze the distributions and functions of the genes.
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