Easier, more reproducible data – starting with the LUMICKS protocols
The single-stranded DNA is shipped as double-stranded DNA that is biotinylated at the 5’ and 5’ ends of the same strand. After melting the DNA one of the strands will fall off and a single stand remains.
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The Protein labeling and tethering kit (ybbR) is a highly efficient kit to study protein dynamics at a single-molecule level through optical tweezers. The methodology is divided into two main procedures: 1. Protein labeling with functionalized DNA handles, 2. Protein tethering to beads for optical tweezers experiments.
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The single-stranded DNA is shipped as double-stranded DNA that is biotinylated at the 5’ and 5’ ends of the same strand. After melting the DNA one of the strands will fall off and a single stand remains.
Click the button below to download the protocol!
The hairpin tethering kit is a highly efficient system to characterize secondary DNA/RNA structures using high-resolution single-molecule optical tweezers.
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When using any of the standard reusable LUMICKS flow cells, it is important to follow robust cleaning protocols between experiments to avoid cross-contamination.
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DNA structural mechanics kit (for training) protocol
Fluorescent intercalators can bind between basepairs and are very suitable to visualize double-stranded DNA.
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The length of the nick-free biotinylated double-stranded λ-DNA (48,502 bp) makes it ideal for assessing different types of DNA binding molecules. Exposing the DNA to fluorescently labeled proteins enables you to see dynamic binding activities and quantify DNA-associated events, such as replication, repair, organization, or transcription.
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To prevent damage to (short) DNA tethers or bleaching of fluorophores by reactive oxygen species, it is recommended to use an oxygen scavenging system. Here we describe two systems, one that couples the glucose oxidase and catalase (GODCAT) in a buffer containing millimolar glucose and the other that employs Trolox. While the two systems can be used separately, a combination of both systems is recommended for optimal results.
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The Protein labeling and tethering kit (Cysteine) is a highly efficient system to study protein dynamics at a single-molecule level through optical tweezers. The methodology is divided into two main procedures: 1. Protein labeling with functionalized DNA handles, 2. Protein tethering to beads for optical tweezers experiments.
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T7 DNA polymerase will either include (polymerization) or exclude (exonucleolysis) basepairs, depending on the force that you apply. This kit allows you to study these kinetics of enzymatic activities associated with DNA replication.
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The hairpin tethering kit is a highly efficient system to characterize secondary DNA/RNA structures using high-resolution single-molecule optical tweezers.
Click the button below to download the protocol!