dynamic HK-EA-1 v1.1 DNA Enzyme Activity Kit with Red Dye Ra User Manual

September 15, 2024
Dynamic

dynamic HK-EA-1 v1.1 DNA Enzyme Activity Kit with Red Dye Ra

dynamic-HK-EA-1-v1-1-DNA-Enzyme-Activity-Kit-with-Red-Dye-Ra-
product

Specifications

  • Product Name: heliX+ DNA Enzyme Activity Kit with red dye Ra
  • Manufacturer: Dynamic Biosensors GmbH & Inc.
  • Model: HK-EA-1 v1.1

Product Description
The heliX+ DNA Enzyme Activity Kit with red dye Ra includes two spots with two different anchor sequences for DNA-encoded addressing. It comes with Adapter strands pre-hybridized to Template-32-P-Ra DNA to facilitate biochip functionalization.

Usage Instructions

Preparation

  1. Mix Adapter strand 1 – nf / Template-32-P-Ra DNA and cTemplate Adapter strand 2 – nf / Template-32-P-Ra DNA at a 1:1 ratio in the same vial.
  2. The solution is now ready for biochip functionalization.

Assay Setup in heliOS
To study enzymatic activity, follow these steps in heliOS:

  1. Create a New Assay Workflow.
  2. Add Custom Assay and load Enzyme Binding and Activity.
  3. Modify parameters based on your needs and run the assay.

Binding Kinetics
Perform enzyme binding kinetics to determine the Kd before running the assay. Use the obtained Kd as the initial test  concentration for enzyme association to avoid overcrowding the surface.

Substrate Scouting:
Choose a broad concentration range for the substrate to obtain reliable sigmoidal fit during extraction of kcat and KM. Use a minimum of 6 concentrations for best results.

FAQ

Q: Can I use this kit for inhibition assay?
A: For inhibition assays or any further questions, please contact the support team at support@dynamicbiosensors.com.

DNA ENZYME ACTIVITY KIT
with red dye Ra
Dynamic Biosensors GmbH & Inc.
HK-EA-1 v1.1

Key Features

  • ssDNA template (48 + 32 bp) for functionalization of heliX® Adapter Chip on Spot 1.
  • dsDNA template (48 + 32 bp) for functionalization of heliX® Adapter Chip on Spot 2.
  • Compatible with heliX® Adapter Chip.
  • Includes Adapter strands for 100 regenerations.
  • This DNA template carry a moderately hydrophilic red dye (Ra) with a single positive net charge.

heliX® Adapter Chip Overview

2 spots with 2 different anchor sequences for DNA-encoded addressing.

dynamic-HK-EA-1-v1-1-DNA-Enzyme-Activity-Kit-with-Red-Dye-
Ra-

Product Description

Order Numbe r: HK-EA-1
Table 1. Contents and Storage Information

Material Cap Concentration Amount Buffer Storage

Adapter strand 1 – nf /

Template-32-P-Ra DNA

| Transparent| 200/250 nM| 10 x 100 µL| TE40 [ 1]| -20°C
cTemplate Adapter strand 2 – nf /

Template-32-P-Ra DNA

| Black| 200/250 nM| 10 x 100 µL| TE40 [ 1]| -20°C

For research use only.
This product has a limited shelf life, please see expiry date on label.
To avoid many freeze thaw cycles please aliquot the nanolever.

Preparation

IMPORTANT
Both Adapter strands are already pre-hybridized. Adapter strand 1 to the Template-32-P-Ra DNA strand, leaving the upper part as ssDNA, and cTemplate- Adapter strand 2 to the Template-32-P-Ra DNA strand, leaving Spot 2 completely as dsDNA.

Next, simply mix in the same vial the Adapter strand 1 – nf / Template-32-P-Ra DNA (200/250 nM) and the cTemplate Adapter strand 2 – nf / Template-32-P-Ra DNA (200/250 nM) at 1:1 ratio (v/v).
Solution is ready to use for biochip functionalization.

Useful Order Numbers

Table 2. Order Numbers

Product Name Comment Order No
heli X ® Adapter Chip Chip with 2 detection spots ADP-48-2-0
10x Passivation solution For passivation of chip surface SOL-PAS-1-5
Regeneration solution For regeneration of chip surface SOL-REG-1-5

Assay Setup in heliOS

For studying enzymatic activity of a nucleic acid modifying enzyme (e.g., polymerase, reverse transcriptase, helicase, etc.).
Go to heliOS > create a New Assay Workflow > add Custom Assay > load Enzyme Binding and Activity > modify the parameters based on your needs and run the assay.
Suggested assay parameters (e.g., flow rates, functionalization time, LED power, etc.) are within the heliOS assay.

TIP
It is strongly recommended to perform binding kinetics of the enzyme beforehand. The obtained Kd during enzyme kinetics can be the initial test concentration for the association of the enzyme during enzyme kinetics. This concentration is a good compromise to not overcrowd the surface and avoid multiple enzymes binding to the same template.

TIP
For an initial scouting of the substrate, choose a broad concentration splitting spanning the low nanomolar to high micromolar, and a blank. A minimum of 6 concentrations of substrate are recommended to obtain a reliable sigmoidal fit during the extraction of the kcat and KM.

For inhibition assay, analysis or any further questions, please contact the support team at support@dynamic-biosensors.com.

Contact

Dynamic Biosensors GmbH
Perchtinger Str. 8/10
81379 Munich
Germany

Dynamic Biosensors, Inc.
300 Trade Center, Suite 1400
Woburn, MA 01801
USA

Order Information order@dynamic-biosensors.com

Technical Support

support@dynamic-biosensors.com

www.dynamic-biosensors.com
Instruments and chips are engineered and manufactured in Germany.
©2024 Dynamic Biosensors GmbH | Dynamic Biosensors, Inc. All rights reserved.

[1] TE40: 10 mM Tris, 40 mM NaCl, 0.05 % Tween20, 50 μM EDTA, 50 μM EGTA
HK-EA-1 v1.1
www.dynamic-biosensors.com

References

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