sparQ Universal Library Quant Kit
- Faster time to results – 50% shorter run time than traditional cycling protocols
- Accurate, reliable quantification of NGS libraries of various sizes and GC-content
- High amplification efficiency across a wide linear dynamic range
- Stabilized, ready-to-use sparQ Universal Fast Mastermix to reduce pipetting steps
- Superior run to run uniformity ensuring highly precise measurements
sparQ Universal Library Quant Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention or treatment of a disease.
sparQ Universal Library Quant Kit
Description
sparQ Universal Library Quant Kit provides rapid and accurate quantification of libraries prepared for sequencing on Illumina® NGS platforms. Accurate quantification of the number of amplifiable library molecules prior to loading onto a flow cell is a critical step in the NGS workflow and it ensures optimal cluster generation and cost-effective use of sequencing capacity. The sparQ Universal Library Quant Kit uses real-time quantitative PCR (qPCR) to specifically quantify the number of library molecules that possess the appropriate adapter tag at each end.
Watch the step-by-step tutorial illustrating the sparQ Universal Library Quant Kit protocol
Details
|
Name |
Description |
Volume (100 rxn kit) |
Volume (500 rxn kit) |
|
sparQ Universal Fast Mastermix |
1.25x sparQ Universal Fast Mastermix containing primers that target the P5 and P7 Illumina adaptor sequences |
1 x 1.6 ml |
1 x 8 ml |
|
sparQ DNA Standard 1 |
20 pM linear, dsDNA standard in 1x Library Dilution Buffer |
1 x 25 µl |
1 x 250 µl |
|
sparQ DNA Standard 2 |
2 pM linear, dsDNA standard in 1x Library Dilution Buffer |
1 x 25 µl |
1 x 250 µl |
|
sparQ DNA Standard 3 |
0.2 pM linear, dsDNA standard in 1x Library dilution buffer |
1 x 25 µl |
1 x 250 µl |
|
sparQ DNA Standard 4 |
0.02 pM linear, dsDNA standard in 1x Library Dilution Buffer |
1 x 25 µl |
1 x 250 µl |
|
sparQ DNA Standard 5 |
0.002 pM linear, dsDNA standard in 1x Library Dilution Buffer |
1 x 25 µl |
1 x 250 µl |
|
sparQ DNA Standard 6 |
0.0002 pM linear, dsDNA standard in 1x Library Dilution Buffer |
1 x 25 µl |
1 x 250 µl |
|
High ROX Reference Dye |
High ROX reference dye for use with “High ROX” instruments. |
1 x 25 µl |
1 x 250 µl |
|
Library Dilution Buffer (10x) |
10x concentrated buffer containing 0.1M Tris-HCl (pH 8.0), 1 mM EDTA, and stabilizers |
1 x 1.5 ml |
5 x 1.5 ml |
Instrument Compatibility
ROX
- Applied Biosystems 5700
- Applied Biosystems 7000
- Applied Biosystems 7300
- Applied Biosystems 7700
- Applied Biosystems 7900
- Applied Biosystems 7900HT
- Applied Biosystems 7900 HT Fast
- Applied Biosystems StepOne™
- Applied Biosystems StepOnePlus™
Low ROX
- Applied Biosystems 7500
- Applied Biosystems 7500 Fast
- Stratagene Mx3000P®
- Stratagene Mx3005P™
- Stratagene Mx4000™
- Applied Biosystems ViiA 7
- Applied Biosystems QuantStudio™ (all models)
- Douglas Scientific IntelliQube®
No ROX
- Quantabio Q
- BioRad CFX
- Roche LightCycler 480
- QIAGEN Rotor-Gene Q
- Agilent AriaMx
- Azure Cielo™
- Analytik Jena qTower
- Analytik Jena qTOWERiris
- Other
Bio-Rad iCycler iQ systems
- BioRad iCycler iQ™
- BioRad MyiQ™
- BioRad iQ™5
Resources
Flyers
Product Manuals
Application Notes
Brochures
CofA (PSFs)
SDSs
Publications
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Customer Product Reviews
| 5 star | 25% | |
| 4 star | 25% | |
| 3 star | 50% | |
| 2 star | 0% | |
| 1 star | 0% |
sparQ Universal Library Quant Kit
Customer Testimonials
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Straightforward and easy to use, while we don’t need to order again currently, it will be on our future purchase list. The only slight improvement is the explanation for using the green colour in the qPCR setup; the Rox aspect is explained in detail, which was a little confusing. And I needed some minutes to figure out the programming of our machine with the explanation. Otherwise, the explanation was beneficial, the program fast, and the calculation helpful.
In our tests, the kit did not perform so well at the low edge of the tested concentrations.
Otherwise it is a fast alternative to our standard choice for large number of samples!
The prep for qPCR is very straightforward, but the results are VERY different than those obtained with 2 other kits (NEB and PCRBio). I obtain values off ~40-50% higher than those from the other kits, so im worried that using this kit would lead to the overestimation of library concentrations. The price is definitely unrivaled (for now), so i may give it another shot with the leftover kit i have.
I would also suggest another improvement – the addition of a blue dye so it’s easier to pipette mixture into PCR wells, especially for small wells, such as those for 384-well plates. PCRBio offers kits with blue dyes and I see no issue with performance.
The product worked fine but we only perform qPCR on a handful (or fewer samples) at one time, so the volume of the standards that come with the kit are insufficient.