4X One-Step RT-qPCR Master Mix, 50 mL (No passive dye)

Cat. No.: LAU-DC-LWMM4X

Size: 50 mL ( 4X 1-Step RT-qPCR Master Mix )

Storage:  -20°C or below

Note: Follow safe chemical handling protocols. Wear appropriate protective eyewear, clothing, and gloves.
Please, Ask for your trial sample

Intended Use and Product Description:

The One-Step RT-qPCR Master Mix was designed for sensitive and reproducible detection of RNA and/or DNA targets in a single multiplex RT-qPCR reaction. The 4X format was developed to provide simple reaction setup and robust performance. The Master Mix has been formulate,(ISO 13485, cGMP compliant facility), ensuring stringent production and quality control measures.

The 4X One-Step Master Mix (No passive dye) is used to perform one-step single or multiplexed real time PCR applications with any gene-specific primer and probe sets and is suitable for both RNA and DNA targets.
Isolate and purify the target nucleic acid samples according to your laboratory practices. Contact Launchworks for sample preparation recommendations.

Avoid excess freeze-thaw cycles on each tube of Master Mix.
Store Master Mix at or below -20˚C per expiration dating on label. Do not heat, incubate, or keep at ambient temperature. Keep mix on ice while setting up RT-PCR assays.

Stand Out Features: Only three steps from sample prep (i.e. RNA extraction) to getting results from your RT-qPCR results:
(1) reagent mixing,  (2) plate dispensing,  (3) RT-qPCR method running.

High-Quality Enzymes:
The master mix is made up of 4 highly specific and active enzymes:
Specifications:

Product Name	4X One-Step RT-qPCR Master Mix (No ROX)
Components	4X mix requiring no passive dye. 1 x 500 uL in a mL microtube (100 reactions per tube), low detergent
Shelf Life	12 months
Storage Temp	-20°C or below
Assay Name/Specification (min release criteria)	4X One-Step RT-qPCR Master Mix (No ROX) is functionally tested in One-Step RTqPCR with SARS CoV-2 E-gene RNA template, resulting in a standard curve across a dynamic range of 6 orders of magnitude
Individual Product Component Testing	Quality Control Tests including activity testing protocols are performed for each component included in the 4X One-Step RT-qPCR Master Mix (No ROX) and meet the designated specifications of RNAse, DNAse and Protease Free

Protocol: Download Protocol

1. Thaw all reagents and keep on ice until set-up is complete.
2.Add the following to one well per sample. Calculate the totalvolume required for each reaction based on the following table.

Component	Standard System Set-Up (20uL/reaction)	Notes
RNAse-Free Water	9 μL	Should be RT-PCR Grade Nuclease-Free Water.
Primer and Probe Set (20X – see note)	1 μL	Recommended final primer concentrations of 400-600nM and a probe concentration of 200-250nM.
1-Step Master Mix(4X)	5 μL	Ensure 4X Master Mix is thoroughly mixed in each well.
RNA or DNA Sample or Control	5 μL	This may be adjusted if needed, by also adjusting the volume of water. Do not exceed 20uL total per reaction.
Total Volume Per Reaction	20 μL	Mix contents of each reaction thoroughly, once all components are present. Centrifugation of plates or tubes for 3 min at 2,000g (4˚C) is recommended to avoid bubbles.

3.RT-real-time PCR Thermal Cycling Conditions:

Step	Cycles	Temperature	Time
Reverse Transcription	1	53 ˚C	Hold 10 mins. (ramp rate 4.4)
Denaturation	1	95 ˚C	Hold 2 mins. (ramp rate 4.4)
Amplification	45	95 ˚C	Hold 15 sec. (ramp rate 4.4)
		60 ˚C	Hold 30 sec. (ramp rate 2.2)
Cooling	1	40 ˚C	Hold 30 sec. (ramp rate 2.2)

Note: Data acquisition during 60˚C hold of Amplification phase. Ramp rates may vary depending on PCR equipment.

Shelf Life:
Internal shelf-life testing is currently ongoing and is currently set at 12 months when stored at or below -20˚C.

Warnings and Limitations:
This material is sold as Research Use Only and should be used in a laboratory setting by trained and qualified professionals only.
While we make every effort to ensure the safety of our products, we recommend handling any biological materials with standard precautions as if capable of spreading infectious disease. We do not recommend use of the product if it has passed its expiration date, or if there is evidence of leakage.