biotechrabbit | 14X LYO-ready CAPITAL™ qPCR Probe Mix

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14X LYO-ready CAPITAL™ qPCR Probe Mix

Features

Freeze-dryable formulation including excipients and stabilizers
Best-in-class performance in both single and multiplex quantitative amplification
Convenient master mix for low-copy number detection
High specificity and sensitivity

Applications

For use with standard and fast qPCR platforms
Single and multiplex qRT-PCR amplification
Compatible with a wide range of probe technologies including
Taqman®, Molecular Beacons® and Scorpion® probes

Purchase 14X LYO-ready CAPITAL™ qPCR Probe Mix

Availability: In stock Bulk requests please inquire at oem@biotechrabbit.com

Catalog #	Size	Package Content	Price*	Qty
BC0504305	500 reactions of 20 µl	750 µl 14X LYO-ready CAPITAL qPCR Probe Master Mix 3 ml 4X qPCR Probe Reconstitution Buffer	€370.00

*Does not include applicable tax, shipping & handling, or other charges.

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biotechrabbit™ 14X LYO-ready CAPITAL™ qPCR Probe Master Mix is a ready-to-use and concentrated formulation of all enzymes, dNTPs, excipients, and stabilizers for producing home-made lyophilizates with the option to include primers and probes. The time-effective lyophilization is possible for high and low volumes of the master mix without the collapse of the lyophilizate. The resulting lyophilizate is highly resistant to humidity and is swiftly dissolved in the provided reconstitution buffer.

The master mix is optimized for real-time PCR quantification of viral DNA, cDNA and genomic DNA from a wide range of targets. The mix ensures high specificity and sensitivity in single and multiplex amplification, making it the best choice for extremely low-copy-number targets in pathogen detection.

CAPITAL qPCR Probe Master Mix uses a proprietary enzyme and buffer formulation suitable for fast extension and multiplexing in challenging PCR reactions.

Component	Composition
14X LYO-ready CAPITAL™ qPCR Probe Master Mix	14X concentrated formulation of a qPCR Probe Master Mix including excipients, ready for lyophilization
4X qPCR Probe Reconstitution Buffer	Optimized PCR reconstitution buffer in 4X concentration

Storage

−20 °C (until expiry date, see product label)

Handling

Prevention of contamination

Contamination with undesired DNA is a concern when assembling the amplification reactions. To eliminate the possibility of contamination with undesired DNA, follow the guidelines below:

Wear disposable gloves when handling the solutions.
Dedicate separate sterile areas for the preparation of samples and reaction mixtures.
Use molecular-grade nuclease-free water and reagents.
Include a non-template control reaction in every PCR assay.
Avoid carryover contamination.

Recommendations before starting a lyophilization project

The formulation is optimized for freeze-drying a minimum of 5 reactions of 20 µl.
Optional: Prepare concentrated primers and probes in molecular biology grade water and restrict the volume to 20 % of the master mix.

Calculation examples

Three calculation examples for 20 µl reaction size

Parameter	Example A	Example B	Example C
qPCR volume	20 µl	20 µl	20 µl
Reactions per bead/cake	5	50	100
Number of beads/cakes	1	10	50
Total number of reactions	5	500	5000
Total reaction volume	100 µl	10 ml	100 ml

Pipetting scheme

Divide the total reaction volume above by a factor of 14. Minimize the volume of primers and probes.

Component	Example A	Example B	Example C
14X LYO-ready CAPITAL™ qPCR Probe Master Mix	7.1 µl	71 µl	7.1 ml
Optional: Primer & Probes (max. 20 % v/v)	1.4 µl	14 µl	1.4 ml
Volume to freeze-dry per bead/cake	8.5 µl	85 µl	170 µl

Suggested lyophilization protocol

Step	Time*	Temperature	Pressure
Freezing	120 min	−40 °C	—
Primary drying	600 min	−40 °C	0.12 mbar / 12 Pa
Secondary drying	300 min	25 °C	0.01 mbar / 1 Pa

*Duration of freeze-drying cycles depends on the lyophilizate volume, the equipment, and the type and dimensions of the vials.

Reconstitution of lyophilized master mix

Add the required volume of the reconstitution buffer to the lyophilized CAPITAL™ qPCR Probe Master Mix.
Mix well. The lyophilizate should dissolve swiftly.
Store the reconstituted CAPITAL™ qPCR Probe Master Mix at −20 °C.

Parameter	Example A	Example B	Example C
4X Reconstitution Buffer per bead/cake	25 µl	250 µl	500 µl
Total volume of 4X Reconstitution Buffer	25 µl	2.5 ml	25 ml

Cycling program

Step	Temperature	Time	Cycles
Initial activation	95 °C	3 min	1
Denaturation	95 °C	10 s	40–45
Annealing/Extension*	60–68 °C*	30 s	40–45

*Recommended annealing/extension temperature is primer Tm +2 °C. Use gradient PCR to optimize the annealing temperature. Do not use temperatures below 60 °C.

Product manual