biotechrabbit | 4X LYO-ready CAPITAL™ 1-Step qRT-PCR Probe Mix Plus

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4X LYO-ready CAPITAL™ 1-Step qRT-PCR Probe Mix Plus

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 pathogen detection
High specificity and sensitivity across a wide range of sample sources
Minimized risk of carryover contamination
Compatible with a wide range of probe technologies, including TaqMan®, Molecular Beacons® and Scorpion® probes

Applications

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

Purchase 4X LYO-ready CAPITAL™ 1-Step qRT-PCR Probe Mix Plus

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

Catalog #	Size	Package Content	Price*	Qty
BR0505301	100 reactions of 20 µl	1 × 500 µl 4X LYO-ready CAPITAL™ 1-Step qRT-PCR Probe Master Mix Plus 1 × 500 µl 4X qRT-PCR Probe Reconstitution Buffer	€92.00
BR0505302	500 reactions of 20 µl	5 × 500 µl 4X LYO-ready CAPITAL™ 1-Step qRT-PCR Probe Master Mix Plus 5 × 500 µl 4X qRT-PCR Probe Reconstitution Buffer	€440.00

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

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biotechrabbit™ 4X LYO-ready CAPITAL™ 1-Step qRT-PCR Probe Master Mix Plus is a ready-to-use, four-times-concentrated formulation containing an adjusted concentration of heat-labile Uracil-DNA Glycosylase (HL-UDG) to minimize the risk of carryover contamination. The mix includes all enzymes, dNTPs, excipients and stabilizers required for producing in-house lyophilizates, with the option to include primers and probes.

Time-efficient lyophilization is possible for both high and low volumes of the master mix without collapse of the lyophilizate. The resulting lyophilizate is highly resistant to humidity and dissolves rapidly in the provided reconstitution buffer. 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. It ensures high specificity and sensitivity in single and multiplex amplification, making it an excellent choice for extremely low-copy-number targets in pathogen detection.

4X LYO-ready CAPITAL™ 1-Step qRT-PCR Probe Master Mix Plus uses a proprietary formulation for efficient reverse transcription and qPCR in a single tube.

Component	Composition
4X CAPITAL™ 1-Step qRT-PCR Probe Master Mix Plus	4X concentrated formulation of a CAPITAL™ 1-Step qRT-PCR Probe Master Mix Plus
4X qRT-PCR Probe Reconstitution Buffer	Optimized qRT-PCR reconstitution buffer in 4X concentration

Storage

Store at −20 °C until expiry date.

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.

Recommendations before starting a lyophilization project

The formulation is optimized for freeze-drying both single and multiple reactions with a reaction volume of 20 µl.
Optional: Prepare concentrated primers and probes in molecular-biology-grade water and restrict their volume to a maximum of 20 % of the LYO-ready master mix.

Calculation examples

Three calculation examples for a 20 µl reaction size:

Parameter	Example A	Example B	Example C
qRT-PCR volume	20 µl	20 µl	20 µl
Reactions per bead/cake	1	5	25
Number of beads/cakes	1	100	200
Total number of reactions	1	500	5000
Total reaction volume	20 µl	10 ml	100 ml

Pipetting scheme

Divide the total reaction volume above by a factor of 4. Minimize the volume of primers and probes. Do not add water.

Component	Example A	Example B	Example C
4X LYO-ready CAPITAL™ 1-Step qRT-PCR Probe Master Mix Plus	5 µl	2.5 ml	25 ml
Optional: Primers and probes (maximum 20 % v/v)	≤1 µl	≤500 µl	≤5 ml
Volume to freeze-dry per bead/cake	5–6 µl	25–30 µl	125–150 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

*The 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 reconstitution buffer to the lyophilized CAPITAL™ 1-Step qRT-PCR Probe Master Mix Plus, as shown in the examples below.
Mix well. The lyophilizate should dissolve rapidly.
Store the reconstituted CAPITAL™ 1-Step qRT-PCR Probe Master Mix Plus at −20 °C.

Parameter	Example A	Example B	Example C
4X Reconstitution Buffer per bead/cake	5 µl	25 µl	125 µl
Total volume of 4X Reconstitution Buffer	5 µl	2.5 ml	25 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

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

Product manual