Detecting protein of transcriptional factor through probe of digestion FRET marke nucleic acid of nuclease
A technology of transcription factors and nucleic acid probes, applied in the field of nuclease digestion of FRET-labeled nucleic acid probes to detect transcription factor proteins, can solve the problems of high chip preparation and use costs
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[0106] Here only FAM / Dabcyl labeled FRET nucleic acid probes were used to detect the transcription factor protein NF- κ The experiment of B is taken as an example to illustrate the specific implementation of the technology of the present invention.
[0107] 1. Design and chemical synthesis of FRET-labeled nucleic acid probes
[0108] We designed and synthesized by BIOASIA Biologic Technology Co.LTD., Shanghai, China, the following two oligonucleotides with reverse complementary base sequences for the preparation of FRET for detecting NF-κB Nucleic acid probe:
[0109] FAM-strand: 5′...AGTTGAG GGGACTTTCC CGAA TCG GGGACTTTCC CAGGC...3'( dT-FAM)
[0110] Dabcyl-strand: 3′...TCAACTC CCCTGAAAGG GCT AAGC CCCTGAAAGG GTCCG...5′( dT-Dabcyl)
[0111] The underlined sequence in the oligonucleotide is the NF-κB binding sequence.
[0112] Dissolve oligonucleotide FAM-strand and Dabcyl-strand in TEN buffer (10mM Tris, 1mM EDTA, 0.1M NaCl, pH8.0) at a concentration of 100...
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