35 nucleotide 5′‐Cy5‐labelled and HPLC‐purified DNA oligonucleotides were purchased from Sigma. To generate dsDNA, equimolar amounts of forward and reverse ssDNA oligonucleotides were mixed in 1xSSC buffer (150 mM NaCl, 15 mM sodium citrate) and incubated for 2 min at 90°C in a Thermomixer. The temperature was reduced to 60°C for 5 min and then further reduced to 20°C for 30 min. For EMSA, 50 nM of nucleic acids was mixed with increasing concentrations of recombinant proteins in a buffer containing 20 mM Tris–HCl pH 8.0, 100 mM KCl, 3mM MgCl2, 1 mM EDTA pH 8.0, 5% glycerol, 0.05% NP‐40, 2 mM DTT, 50 ng/ml yeast tRNA (Thermo Fisher) and 2.5 ng/ml BSA (NEB). Samples were incubated at 4°C with rotation for 1 h and resolved on a 4% polyacrylamide (60:1) gel (25 mM Tris–HCl, 200 mM glycine, 5% glycerol, 0.075% APS, 0.05% TEMED) in 12.5 mM Tris–HCl and 100 mM glycine. The Cy5 signal was scanned on a Typhoon FLA 9500 fluorescence scanner.
35 nucleotide 5 cy5 labelled and hplc purified dna oligonucleotides
Manufactured by Merck Group
35 nucleotide 5′‐Cy5‐labelled and HPLC‐purified DNA oligonucleotides.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using 35 nucleotide 5 cy5 labelled and hplc purified dna oligonucleotides
1
Electrophoretic Mobility Shift Assay for DNA-Protein Interactions
2
Electrophoretic Mobility Shift Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
35 nucleotide 5'-Cy5 labelled and HPLC-purified DNA oligonucleotides were purchased from Sigma. To generate dsDNA, equimolar amounts of forward and reverse ssDNA oligonucleotides were mixed in 1xSSC buffer (150 mM NaCl, 15 mM sodium citrate) and incubated for 2 minutes at 90°C in a Thermomixer. The temperature was reduced to 60°C for 5 minutes, then further reduced to 20°C for 30 minutes. For EMSA, 50 nM of nucleic acids were mixed with increasing concentrations of recombinant proteins in a buffer containing 20 mM Tris-HCl pH 8.0, 100 mM KCl, 3mM MgCl2, 1 mM EDTA pH 8.0, 5% glycerol, 0.05% NP40, 2 mM DTT, 50 ng/ml yeast tRNA (ThermoFisher) and 2.5 ng/ml BSA (NEB). Samples were incubated at 4°C with rotation for 1 hour and resolved on a 4% polyacrylamide (60:1) gel (25 mM Tris-HCl, 200 mM glycine, 5% glycerol, 0.075% APS, 0.05% TEMED) in 12.5 mM Tris-HCl and 100 mM glycine. The Cy5 signal was scanned on a Typhoon FLA 9500 fluorescence scanner.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!