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10X LYO-ready CAPITAL™ 1-Step qRT-PCR 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

One-step amplification of mRNA, total RNA and viral RNA targets
For use with standard and fast qPCR platforms
Single and multiplex qRT-PCR amplification

Purchase 10X LYO-ready CAPITAL™ 1-Step qRT-PCR Probe Mix

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

Catalog #	Size	Package Content	Price*
BR0502901	500 reactions of 20 µl	10X LYO-ready CAPITAL 1-Step qRT-PCR Probe Master Mix 5X qRT-PCR Probe Reconstitution Buffer	€445.00
BR0502903	5000 reactions of 20 µl	10X LYO-ready CAPITAL 1-Step qRT-PCR Probe Master Mix 5X qRT-PCR Probe Reconstitution Buffer	€3,560.00
BR0503001	500 reactions of 20 µl	10X LYO-ready CAPITAL 1-Step qRT-PCR Probe Master Mix 2X qRT-PCR Probe Reconstitution Buffer	€445.00
BR0503003	5000 reactions of 20 µl	10X LYO-ready CAPITAL 1-Step qRT-PCR Probe Master Mix 2X qRT-PCR Probe Reconstitution Buffer	€3,560.00

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

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biotechrabbit™ 10X LYO-ready CAPITAL™ 1-Step qRT-PCR Probe 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 reconstitution buffer is available in two different concentrations.

The master mix is optimized for real-time PCR quantification of RNA templates, including mRNA, total RNA, and viral RNA 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 qRT-PCR Probe Mix uses proprietary formulation for efficient RNA transcription and qPCR in a single tube.

Component	Composition
10X LYO-ready CAPITAL™ qRT-PCR Probe Master Mix	A lyo-ready 10X concentrate including all enzymes, dNTPs, stabilizers, and excipients required for lyophilization.
2X or 5X qRT-PCR Probe Reconstitution Buffer	Proprietary 5X buffer for reconstitution.

Storage

−20 °C (until expiry date)

Handling

Prevention of contamination

RNase contamination is a concern when handling RNA. To minimize the risk of contamination and prevent RNA degradation, 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
qRT-PCR 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 10. Minimize the volume of primers and probes.

Component	Example A	Example B	Example C
10X LYO-ready CAPITAL™ 1-Step qRT-PCR Probe Mix	10 µl	1 ml	10 ml
Optional: Primer & Probes (max. 20 % v/v)	2 µl	200 µl	2 ml
Volume to freeze-dry per bead/cake	12 µl	120 µl	240 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™ 1-Step qRT-PCR Probe Mix.
Mix well. The lyophilizate should dissolve swiftly.
Store the reconstituted CAPITAL™ 1-Step qRT-PCR Probe Mix at −20 °C.

Parameter	Example A	Example B	Example C
2X Reconstitution Buffer per bead/cake	50 µl	500 µl	1 ml
5X Reconstitution Buffer per bead/cake	20 µl	200 µl	400 µl
Total volume of 2X Reconstitution Buffer	50 µl	5 ml	50 ml
Total volume of 5X Reconstitution Buffer	20 µl	2 ml	20 ml

Cycling program

Step	Temperature	Time	Cycles
Reverse transcription	50 °C	10 min	1
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