First, biotin label was linked to the 3′ end of artificially synthesized single-stranded oligonucleotide probe containing binding site by DNA 3′ end biotin label kit (Beyotime, Shanghai, China). Second, double-stranded DNA probe with biotin label was obtained by annealing with artificially synthesized complimentary chain. Third, purified CkREV protein was incubated with probe with biotin label at a certain proportion while unlabeled double-stranded probe was used as cold probe. Fourth, native-PAGE was employed to separate samples before being transferred onto nylon membrane (Solarbio, Beijing, China) with positive charge through wet transformation method. Fifth, the nylon membrane was placed under ultraviolet crosslinker purple (UVP, Upland, USA) at 254 nm, 120 mJ/cm2 for 60 s. Last, colour development was employed on a completely cross-linked nylon membrane by EMSA chemiluminescence kit (Beyotime, Shanghai, China) for observation under chemiluminescence imager. Detailed steps complied with instructions of EMSA chemiluminescence kit (Beyotime, Shanghai, China).
Dna 3 end biotin label kit
Manufactured by Beyotime
Sourced in China
The DNA 3' end biotin label kit is a laboratory product designed to label the 3' end of DNA molecules with biotin. This kit provides the necessary reagents and protocols to facilitate the biotin labeling process. The core function of this kit is to enable the attachment of biotin, a small molecule, to the 3' terminus of DNA strands, which can be useful in various DNA-based applications and analyses.
Automatically generated - may contain errors
2 protocols using dna 3 end biotin label kit
1
Biotin-Labeled Oligonucleotide Probe for Protein Binding
2
Biotin-Labeled Oligo EMSA Protocol
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First, biotin label was linked to the 3' end of arti cially synthesized single-stranded oligonucleotide probe containing binding site by DNA 3' end biotin label kit (Beyotime, China). Second, double-stranded DNA probe with biotin label was obtained by annealing with arti cially synthesized complimentary chain. Third, puri ed CkREV protein was incubated with probe with biotin label at a certain proportion while unlabeled double-stranded probe was used as cold probe. Fourth, native-PAGE was employed to separate samples before transferred onto nylon membrane (Solarbio, China) with positive charge through wet transformation method. Fifth, the nylon membrane was placed under ultraviolet cross linker purple (UVP, USA) at 254 nm, 120 mJ/cm 2 for 60 s. Last, colour development was employed on completely crosslinked nylon membrane by EMSA chemiluminescence kit (Beyotime, China) for observation under chemiluminescence imager. Detailed steps complied with instructions of EMSA chemiluminescence kit from Beyotime Company.
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