DNA / PCR

qPCR Probe LyoBeads

 

are ready-to-use freeze-dried master mixes that contain all components necessary for a rapid, sensitive and reproducible detection and quantification of DNA and cDNA. qPCR Probe LyoBeads are shipped and stored simply at room-temperature in a dry place. This provides a more cost efficient and ecological distribution compared to other master mixes. All necessary components are already included in one LyoBead: an engineered DNA polymerase, an optimized reaction buffer and ultrapure dNTPs. Only primers and probes need to be added. A hot-start formulation of the included DNA polymerase prevents false amplification during the reaction set-up.

 

Article Product Size Price
#2201S qPCR Probe LyoBeads pre-dispensed 1 x 96 well-plate ¬†‚ā¨ 100,-
#2201M qPCR Probe LyoBeads pre-dispensed 5 x 96 well-plate ¬†‚ā¨ 450,-
#2201C customized qPCR Probe LyoBeads already containing target specific primers&probes - Please inquire

Bulk offer: Should you require larger amounts (>1500 beads), significant discounts are available. Customized qPCR Probe LyoBead production and pre-distribution will be provided on demand as well (further information on possible pre-distributing formates please read the detailed product description below). Please contact us to receive an individual quotation.

 

download-pdf Product Manual (PDF)Order

 

 

Example Results

Example Results

Specific quantification over a broad dynamic range.

 

TaqMan-Assay for parvovirus B19, triple determination on a respective plasmid dilution series. Tests were carried out on 109-1 copies/rxn.

More example results from our external collaboration partner can be downloaded herepdf-file in german orin english. 

 

 

Reaction Setup

Reaction Setup

 

Component Volume
LyoBead 0.75 ¬Ķl¬†(volume of one LyoBead)
Primers & Probes x ¬Ķl
Template/Sample extract y¬†¬Ķl
Water fill up to 20 ¬Ķl total reaction volume

 

For non-dispensed LyoBeads, aliquote them with a metallic tweezer into individual reaction tubes or wells as it is shown in this short video.

For already pre-dispensed Lyobeads, briefly tap-down or gently spin down before opening the reaction tubes.

Add primers and probes as well as template/sample extract. Close the PCR tubes, vortex or mix briefly, spin down and start the PCR.

 

 

Applications

Applications

  • Multiplex PCR
  • Realtime PCR
  • Probe-based PCR
  • Pathogene detection
  • qPCRs (Probe-based)
  • Primer extensions¬†
  • Endpoint PCRs
  • Screening / High-throughput PCRs

Typical PCR protocol

Typical PCR protocol

Step Temperature Time
Initial denaturation 95¬įC 2 min
Denaturation 95¬įC 15 sec
Annealing* 54-72¬įC 30 sec¬† ¬† ¬†(25-40 cycles)
Extension 72¬†¬įC 1 min / 1000 bp
Hold <10¬įC hold

*Typically, the annealing temperature is about 3-5¬įC below the calculated melting temperature of the primers used.¬†

 

Detailed product description

 

Pre-dispensed qPCR Probe LyoBeads

 

Each qPCR Probe LyoBead is equivalent to a 20 ¬Ķl PCR reaction and can either be provided pre-dispensed on demand or non-dispensed. qPCR Probe LyoBeads can be pre-dispensed in¬† 8- or 12-strip PCR tubes (upper image), single PCR tubes or in 96-well plates (as you can see in this video here). Non-dispensed qPCR Probe LyoBeads are provided in micro tubes with screw cap (as shown in the lower image).

 

 

Handling of non-dispensed qPCR Probe LyoBeads

 

It is recommended to use a metallic tweezer to aliquote the qPCR Probe LyoBeads as shown in this short video.

 
 

Quality Control

qPCR Probe LyoBeads are tested in real-time qPCR. The product demonstrates linearity of amplification over a specified serial dilution of human genomic DNA. DNA polymerase activity: DNA polymerase activity has been monitored and adjusted to a specific DNA polymerase activity using an artificial DNA template and a DNA primer. Enzyme-concentration has been determined by proteinspecific staining. Please inquire more information at info@mypols.de for the lot-specific concentration. No contamination has been detected in standard test reactions.

 

Storage

Costly cooling chains and storage at -20¬įC are no longer necessary: ¬†qPCR Probe LyoBeads are¬†shipped and stored simply at room-temperature in a dry place. Once opened, the surrounding air humidity is ideally less than 30%. ¬†

 

Composition / Content of the Product

qPCR Probe LyoBeads contain all the components necessary for PCR, including a hot-start formulated Taq DNA polymerase and an optimized buffer including ultrapure dNTPs.

 

 

img-faqsFAQ`s

Which DNA polymerase is included in these LyoBeads?

 

The LyoBeads contain our best performing, hot-start formulated, engineered Taq DNA polymerase. We have chosen this DNA polymerase as it is best suited for multiplex PCRs, pathogen detection qPCRs and all kinds of probe-based qPCRs.

The enzyme's fidelity is comparable with the wild-type Taq DNA polymerase as described in the reference below. 

Fidelity of DNA synthesis by the Thermus aquaticus DNA polymerase.
Biochemistry 1988; 27(16): 6008‚Äď6013. K. R. Tindall, T. A. Kunkel.

 

 

How can I avoid electro static charge of the LyoBeads?

 

We recommend to use metallic tweezers for distributing the LyoBeads into the reaction-tubes. Also anti-static equipment and gloves may prevent electro static problems. It also helps to stand the PCR tubes or plates on a metallic surface.

 

Can you also produce LyobBeads containing other DNA polymerases?

 

Yes we can. Currently we are working on qPCR LyoBeads containing GreenDye and RT-qPCR Beads containing a Volcano2G mutant for you. On request we can also discuss the production of other LyoBeads with you. Please contact us should you be interested!

 

 

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