C-Trap Reference kit

$695

SKU: 00033 Category:

The C-Trap Reference Kit is designed to support user training, fast-track assay troubleshooting, and verify and monitor instrument performance. The kit provides standardized and validated reagents for a complete DNA-protein binding assay, including data analysis.

The C-Trap Reference Kit is best used in combination with a C-Trap with 565 nm and 638 nm lasers.

This kit includes an ATTO 565-labeled version of the transcription factor lac repressor (LacI-ATTO 565) and a biotinylated DNA construct containing 12 repeats of the LacI DNA target sequence, known as the lac operator (LacO). The LacO DNA construct (37.8 kb) is flanked by two ATTO 647N fluorophores, which facilitate locating the optimal focal plane of the DNA tether prior to incubation with LacI-ATTO 565. This DNA construct was assembled using the LUMICKS DNA tethering kit following LUMICKS Golden Gate Assembly Protocol.

With this protocol, the LacO DNA construct is tethered between two streptavidin-coated beads in the C-Trap. The user can visualize and track the binding of LacI-ATTO 565 to the LacO DNA construct over time to create kymographs, which are then analyzed to determine the association (ON) and dissociation (OFF) rates of LacI.

In addition to LacI-ATTO 565 and biotinylated LacO DNA, the kit includes the 10x binding buffer, streptavidin-coated beads (⌀ 4–4.9 µm) and the alkaline cleaning reagent (ACR).

The kit enables 10 measurements. 

Estimated 1-4 weeks delivery time.

Specifications

  • LacI-ATTO 565: 20 μl | ≥2 μM. Store at -20°C Reference Kit.
    Using 2 μl in 250 μl 1x binding buffer is expected to result in an ON rate >0.1 events per second (at least 1 binding event every 10 seconds).

  • Biotinylated LacO DNA (37.8 kb): 25 μl | 60 ng/μl. Store at 4°C.
    Using 2.5 μl in 500 μl of 1x binding buffer is expected to result in fast tethering in the C-Trap.

  • 10x Binding Buffer: 25 ml. Store at RT.
    Composition: 0.4 M Tris-HCl, 0.5 M NaCl, 20 mM MgCl2, 3 mM NaN3 (pH 7.5 @ 25°C).

  • Streptavidin-coated polystyrene beads (⌀ 4–4.9 µm): 50 μl | 0.5% (w/v) (typical dilution factor 1:100). Store at 4°C.
    Storage buffer in PBS with 3 mM sodium azide. Each batch has a specific bead diameter (e.g., 4.5 μm), which falls within the given range of 4.0 to 4.9 μm.
    Using 5 μl in 500 μl of 1x binding buffer is expected to result in fast bead trapping in the C-Trap.
  • 50x Alkaline cleaning reagent (ACR): 3 ml. Store at RT.
    Use as described in the protocol. ⚠️Hazardous material: Handle with care according to the instructions on the label. The Safety Data Sheet (SDS) is available upon request.

Product resources

Protocols

  • Download the full protocol the C-Trap Reference Kit.

  • A quick protocol guide is also available for faster reference

  • Explore our Golden Gate Assembly application note to efficiently ligate multiple DNA fragments into one biotinylated DNA construct for tethering in the C-Trap.

  • Access a detailed step-by-step Golden Gate protocol to assemble DNA constructs with multiple sequence elements. This protocol was used to generate Biotinylated LacO DNA (37.8 kb) with 12 LacO sequence repeats, which is provided with the kit.

DNA Sequence

Download the DNA sequence of the LacO DNA construct. 

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