Home Page/Product Center

dsDNA HS Assay Kit for Qubit®

Product SKU

Specifications

Price

12640ES60

100 rxn

$105.92

12640ES76

500 rxn

$416.26

Introduction

dsDNA HS (high sensitive) Assay Kit for Qubit® is a fast, sensitive and accurate double-stranded DNA (dsDNA) fluorescence quantitative detection kit. The kits include concentrated assay reagent, dilution buffer, and prediluted DNA standards. This kit is highly selective for dsDNA over RNA and is accurate for initial sample concentration from 10 pg/μL to 100 ng/μL. It is an ideal kit for DNA sample quantification (such as NGS input DNA quantification, DNA library quantification). Common contaminants such as salts, free nucleotides, solvents, detergents, or protein are well tolerated in the assay.

Contents

Number

Material

Concentration

12640ES60

12640ES76

12640-A

dsDNA Reagent

200×concentrate in DMSO

250 μL

1.25 mL

12640-B

dsDNA Buffer

Not applicable

50 mL

250 mL

12640-C

dsDNA Standard 1

0 ng/μL in TE buffer

1 mL

5×1 mL

12640-D

dsDNA Standard 2

10 ng/μL in TE buffer

1 mL

5×1 mL

Storage

All the components can be stored at 2-8℃ for one year.

Protocols

1. The operation flow of Qubit

1. Preparation

1)Equilibrate the components in the kit to room temperature before use.

2)Set up the required number of 0.5-mL tubes for standards and samples. Do not label the side of the tube as this could interfere with the sample read. Label the lid of each standard tube correctly. Use only thin-wall, clear, 0.5-mL PCR tubes. Acceptable tubes include Qubit® assay tubes (Cat. No. Q32856) or Axygen® PCR-05-C tubes (Cat. No. 10011-830).

2. Prepare the working solution

Prepare the working solution by diluting the dsDNA Reagent 1:200 in dsDNA Buffer. Use a clean plastic tube each time you prepare working solution. Do not mix the working solution in a glass container. Working solution is prepared for immediate use within 3 hous.

3. Prepare the sample to be tested

1)Add 10 µL of each dsDNA standard to the appropriate tube, then add working solution to individual assay tubes so that the final volume in each tube after adding sample is 200 µL. mix by vortexing 2–3 seconds. Be careful not to create bubbles.

2)Add 1-20 µL of each DNA samples or library to the appropriate tube, then add working solution to individual assay tubes so that the final volume in each tube after adding sample is 200 µL. mix by vortexing 2–3 seconds. Be careful not to create bubbles.

4. Fluorescence Detection

1)Incubate the tube protect from light at room temperature for 2 mins.

2)According to the operating instructions of the Qubit® Fluorometer, select dsDNA High Sensitivity as the assay type.

Appendix

Table Effect of contaminants in the dsDNA HS Assay Kit for Qubit®

Contaminant

Concentration in 10-μL sample

Final concentration in the assay

Result

Salts

Ammonium acetate

200 mM

10 mM

OK

Sodium acetate

200 mM

10 mM

OK

Sodium chloride

200 mM

10 mM

OK

Magnesium chloride

40 mM

2 mM

OK

Organic Solvents

Phenol

2%

0.1%

OK

Ethanol

20%

1%

OK

Chloroform

4%

0.2%

OK

Detergents

Sodium dodecyl sulfate

0.2%

0.01%

OK

Triton X-100

0.02%

0.001%

OK

Other Compounds

Bovine serum albumin

400 μg/mL

20 μg/mL

OK

RNA

1×*

1×*

OK

dNTPs

2 mM

100 μM

OK

Polyethylene glycol

20%

1%

OK

Agarose

2%

0.1%

OK

*1× indicates a concentration equal to the concentration of dsDNA.

15221439726

15221439726