Vesicle free RNA was eliminated by a combined treatment of Protease K (The final concentration 100ug/ml) and RNase A (The final concentration 10ug/ml). First, sEVs enriched fractions were treated with Protease K (TIANGEN, cat. No. RT403) for 30min at 37°C, then treated with RNase A (TIANGEN, cat. No. RT405) for 15min at 37°C.
Protease k
Manufactured by Tiangen Biotech
Sourced in China
Protease K is a serine protease enzyme that is commonly used in molecular biology and biotechnology applications. It is capable of hydrolyzing a wide variety of proteins by cleaving peptide bonds. Protease K is known for its high activity and broad substrate specificity.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using protease k
1
Vesicle-free RNA Elimination Protocol
2
Exosomal miRNA Expression Analysis
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Exosomes were digested by degrading enzymes and detergent. Firstly, a combined treatment of protease K (TIANGEN, China) and RNase A (TIANGEN, China) was added to the exosome preparation. Exosomes were treated with protease K (final concentration 100 µg/mL) for 30 min at 37 °C, then treated with RNase A (final concentration 100 µg/mL) for 15 min at 37 °C. On the other side, exosomal RNA was digested with Tritonx-100 (final concentration 10%) for 30 min at room temperature (RT), followed by RNase A (final concentration 100 µg/mL) for 15 min at 37 °C. The untreated aliquot was served as a control. Exosomal total RNA was extracted to evaluate the expression level of miR-670-3p by qRT-PCR.
3
Extracting Genomic DNA from Food Samples
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Wizard Magnetic DNA Purification System for Food that used to extract genomic DNA was obtained from Promega Corporation; Protease K was obtained from Tiangen Biotech Co., Ltd. The primers and the specific probe based on the fliC gene were designed by Oligo 6.0 software (Table 1) and prepared by Sangon Biotech; the rRAA kit was supplied by Jiangsu Qitian Gene Technology Co. Ltd.
The fluorescence characterization analysis of PMAxx was photographed by confocal laser scanning microscope (CLSM; Leica TCS SP8); the QT-RAA-F1620 detection system was supplied by Jiangsu Qitian Gene Technology Co. Ltd.
The PMAxx (Bosunlife Co. Ltd.) was deliquesced in sterile water to prepare stock solution at the concentration of 20 mM and stored at -20°C in the dark until used; 2.7 mM potassium chloride, 10 mM disodium hydrogen phosphate, 137 mM sodium chloride, and 2 mM potassium dihydrogen phosphate were mixed to prepare PBS, and then diluted to prepare a 0.01 M PBS solution at pH of 7.4.
The fluorescence characterization analysis of PMAxx was photographed by confocal laser scanning microscope (CLSM; Leica TCS SP8); the QT-RAA-F1620 detection system was supplied by Jiangsu Qitian Gene Technology Co. Ltd.
The PMAxx (Bosunlife Co. Ltd.) was deliquesced in sterile water to prepare stock solution at the concentration of 20 mM and stored at -20°C in the dark until used; 2.7 mM potassium chloride, 10 mM disodium hydrogen phosphate, 137 mM sodium chloride, and 2 mM potassium dihydrogen phosphate were mixed to prepare PBS, and then diluted to prepare a 0.01 M PBS solution at pH of 7.4.
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