A kind of rna pulldown method and kit
A technology of RNA probes and magnetic beads, applied in biological testing, material inspection products, measuring devices, etc., can solve the problems of nuclease contamination, low binding specificity between RNA probes and proteins, false positives, etc., and achieve Prevent contamination, save experimental cost, and improve specificity
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[0037] The present invention is illustrated by taking the pulldown effect of the lincRNA HOTAIR probe on the LSD1 protein as an example, and the probe is labeled with desthiobiotin 8-oxoG. The design and labeling methods of probes are well known to those skilled in the art and will not be repeated here. Streptavidin coupled to magnetic beads (BeaverBeads TM Streptavidin, Beaver Biotechnology Co., Ltd.) and affinity binding with dethiobiotin; the whole cell protein is incubated with magnetic beads-RNA probe, and the protein molecule can be specifically combined with the probe; after washing, the non-specific binding protein Molecule removal; finally, elution is performed with an eluent (for streptavidin) to obtain the target probe-protein complex, and then the protein type is identified by Western Blot. Specific steps are as follows:
[0038] S1. Magnetic bead pretreatment
[0039] ① Take 80 μL of streptavidin-containing magnetic beads and place them in an RNase-free EP tube...
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