RNA pulldown assay was conducted to examine the binding of GPRIN1 and miR-654-5p. After lysing, GC cells were treated with RNase-free DNase I (Beyotime). Afterwards, the biotin-labeled wild-type miR-654-5p (bio-miR-654-5p WT), biotin-labeled mutant miR-654-5p (bio-miR-654-5p MUT) or bio-NC, and streptavidin-coated magnetic beads (Sigma-Aldrich, St Louis, MO, USA) were incubated with cell lysate at 4°C for 3 h. Finally, RNA samples were isolated by TRIzol and subjected to RT-qPCR analysis.
Rnase free dnase 1
Manufactured by Beyotime
RNase-free DNase I is an enzyme used in molecular biology laboratories to selectively degrade DNA while leaving RNA intact. It is commonly used in sample preparation and purification procedures to remove genomic DNA.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using rnase free dnase 1
1
GPRIN1-miR-654-5p Binding Evaluation
2
Verifying KCNMB2-AS1 and miR-3194-3p Binding
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RNA pulldown assay was carried out as previously described [38 (link)] to verify the binding capacity between KCNMB2-AS1 (or PYGL) and miR-3194-3p. After lysing, ESCA cells were treated with RNase-free DNase I (Beyotime). Afterward, the biotin-labeled wild-type miR-3194-3p (bio-miR-3194-3p Wt) or bio-NC and streptavidin-coated magnetic beads (Thermo Fisher Scientific) were incubated with cell lysate at 4°C for 3 h. Finally, RNA samples were purified by TRIzol for RT-qPCR analysis.
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