12 nM Cy3 conjugated primer-template (RP25Cy3/RP266), pssDNA (RP348Cy3/RP343P), or ssDNA (RP348Cy3) was mixed with the indicated amounts of Polθ WT or Polθ L2 in 20 µl of buffer (25 mM Tris-HCl pH 7.5, 30 mM NaCl, 0.01% NP-40, 1 mM DTT, 0.1 mg/ml BSA, 0.5 mM MgCl and 10% glycerol) on ice (primer-template) or at room temp (pssDNA) for 60 min. Reactions were resolved in non-denaturing 5% polyacrylamide gels containing 2.5% glycerol and DNA was visualized using a MultiImage III fluorescence imager (Alpha Innotech) and FluorChem Q software by monitoring Cy3 fluorescence.
Multiimage 3 fluorescence imager
Manufactured by Bio-Techne
The MultiImage III is a fluorescence imager designed for imaging a variety of fluorescent samples. It offers multi-channel detection capabilities and can capture high-resolution images of gels, blots, and other fluorescent-based samples.
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Lab products found in correlation
2 protocols using multiimage 3 fluorescence imager
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Polθ Binding to DNA Substrates
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Polθ Binding to DNA Substrates
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12 nM Cy3 conjugated primer-template (RP25Cy3/RP266), pssDNA (RP348Cy3/RP343P), or ssDNA (RP348Cy3) was mixed with the indicated amounts of Polθ WT or Polθ L2 in 20 µl of buffer (25 mM Tris-HCl pH 7.5, 30 mM NaCl, 0.01% NP-40, 1 mM DTT, 0.1 mg/ml BSA, 0.5 mM MgCl and 10% glycerol) on ice (primer-template) or at room temp (pssDNA) for 60 min. Reactions were resolved in non-denaturing 5% polyacrylamide gels containing 2.5% glycerol and DNA was visualized using a MultiImage III fluorescence imager (Alpha Innotech) and FluorChem Q software by monitoring Cy3 fluorescence.
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