UltraPlex 1-Step ToughMix

Product	Kit Size	Order Info
UltraPlex 1-Step ToughMix	100 x 20 µL rxns (1 x 500 µL)	Order (95166-100)
	500 x 20 µL rxns (5 x 500 µL)	Order (95166-500)
	1000 x 20 µL rxns (1 x 5 mL)	Order (95166-01K)
UltraPlex 1-Step ToughMix ROX	100 x 20 µL rxns (1 x 500 µL)	Order (95167-100)
UltraPlex 1-Step ToughMix Low ROX	100 x 20 µL rxns (1 x 500 µL)	Order (95168-100)
	500 x 20 µL rxns (5 x 500 µL)	Order (95168-500)

Documents

Categories: Brands, QuantaBio

Description

Description

Tough-tested, multiplex, 1-step real time PCR

Features & Benefits

4X concentrated master mix allows greater flexibility with dilute samples – as little as 1 pg of total RNA
Multiplexing capable with up to 5 targets
Tough-tested for applications where PCR inhibition from template impurities may be a concern, such as: crude extracts, clinical specimens, or environmental samples.
Temperature stabilized master mix enables convenient reaction setup at ambient room temperature

UltraPlex 1-Step ToughMix is intended for molecular biology applications. This product is not intended for the diagnosis, prevention or treatment of a disease.

Description

UltraPlex 1-Step ToughMix is a ready-to-use, 4X concentrated master mix 1-step reverse transcription and real-time quantitative PCR (RT-qPCR) of RNA templates using probe-based detection methods. First-strand cDNA synthesis and PCR amplification are carried out in the same tube without opening between steps. Optimized to deliver highly sensitive quantification of RNA viruses or low abundance RNA targets in uni- or highly multiplexed RNA detection assays, this reagent chemistry is optimized to deliver maximum assay sensitivity, precision and reproducibility with miniaturized reaction volumes and either conventional or accelerated thermal cycling protocols. UltraPlex 1-Step ToughMix contains all required components for RT-qPCR except RNA template and probe and is compatible with all dual-labeled probe chemistries.

UltraPlex 1-Step ToughMix (Low ROX/ ROX) (4X), contains:

4X reaction buffer containing optimized titrations of qPCR-grade dATP, dCTP, dGTP, dTTP, and magnesium.
Ultra-pure recombinant proteins: qScript XLT reverse transcriptase, RNase inhibitor protein, and AccuStart II hot-start Taq DNA polymerase.
Proprietary stabilizers additives and optimized ROX passive reference dye (if applicable).

Storage & Handling

Store components in a constant temperature freezer at -25°C to -15°C protected from light upon receipt. For lot specific expiry date, refer to package label, Certificate of Analysis or Product Specification Form.
Repeated freezing and thawing does not affect RT-qPCR performance.

Instrument Capability

ROX

Applied Biosystems 5700
Applied Biosystems 7000
Applied Biosystems 7300
Applied Biosystems 7700
Applied Biosystems 7900
Applied Biosystems 7900HT
Applied Biosystems 7900 HT Fast
Applied Biosystems StepOne™
Applied Biosystems StepOnePlus™

Low ROX

Applied Biosystems 7500
Applied Biosystems 7500 Fast
Stratagene Mx3000P®
Stratagene Mx3005P™
Stratagene Mx4000™
Applied Biosystems ViiA 7
Applied Biosystems QuantStudio™
Agilent AriaMx
Douglas Scientific IntelliQube®

No ROX

Quantabio Q
BioRad CFX
Roche LightCycler 480
QIAGEN Rotor-Gene Q
Other

Bio-Rad iCycler iQ systems

BioRad iCycler iQ™
BioRad MyiQ™
BioRad iQ™5

Related Resources

Benjamin Lee, Vaccine – 2020

ABSTRACT
Background Oral, live-attenuated rotavirus vaccines suffer from impaired immunogenicity and efficacy in low-income countries. Increasing the inoculum of vaccine might improve vaccine response, but this approach has been inadequately explored in low-income countries. Methods We performed a double-blind, parallel group, randomized controlled trial from June 2017 through June 2018 in the urban Mirpur slum of Dhaka, Bangladesh to compare vaccine take (primary outcome) among healthy infants randomized to receive either the standard dose or double the standard dose of oral Rotarix (GlaxoSmithKline) vaccine at 6 and 10 weeks of life. Infants with congenital malformations, birth or enrollment weight <2000 gm, known immunocompromising condition, enrollment in another vaccine trial, or other household member enrolled in the study were excluded. Infants were randomized using random permuted blocks. Vaccine take was defined as detection of post-vaccination fecal vaccine shedding by real-time reverse transcription polymerase chain reaction with sequence confirmation or plasma rotavirus-specific immunoglobulin A (RV-IgA) seroconversion 4 weeks following the second dose. Results 220 infants were enrolled and randomized (110 per group). 97 standard-dose and 92 high-dose infants completed the study per-protocol. For the primary outcome, no significant difference was observed between groups: vaccine take occurred in 62 (67%) high-dose infants versus 69 (71%) standard-dose infants (RR 0.92, 95% CI 0.67–1.24). However, in post-hoc analysis, children with confirmed vaccine replication had significantly increased RV-IgA responses, independent of the intervention. No significant adverse events related to study participation were detected. Conclusions Administration of double the standard dose of an oral, live-attenuated rotavirus vaccine (Rotarix) did not improve vaccine take among infants in urban Dhaka, Bangladesh. However, improved immunogenicity in children with vaccine replication irrespective of initial inoculum provides further evidence for the need to promote in-host replication and improved gut health to improve oral vaccine response in low-income settings. ClinicalTrials.gov: NCT02992197.

The effect of increased inoculum on oral rotavirus vaccine take amonginfants in Dhaka, Bangladesh: A double-blind, parallel group,randomized, controlled trial

Benjamin Lee, Vaccine – 2020

ABSTRACT
Background:Oral, live-attenuated rotavirus vaccines suffer from impaired immunogenicity and efficacyin low-income countries. Increasing the inoculum of vaccine might improve vaccine response, but thisapproach has been inadequately explored in low-income countries.Methods:We performed a double-blind, parallel group, randomized controlled trial from June 2017through June 2018 in the urban Mirpur slum of Dhaka, Bangladesh to compare vaccine take (primary out-come) among healthy infants randomized to receive either the standard dose or double the standard doseof oral Rotarix (GlaxoSmithKline) vaccine at 6 and 10 weeks of life. Infants with congenital malforma-tions, birth or enrollment weight <2000 gm, known immunocompromising condition, enrollment inanother vaccine trial, or other household member enrolled in the study were excluded. Infants were ran-domized using random permuted blocks. Vaccine take was defined as detection of post-vaccination fecalvaccine shedding by real-time reverse transcription polymerase chain reaction with sequence confirma-tion or plasma rotavirus-specific immunoglobulin A (RV-IgA) seroconversion 4 weeks following the sec-ond dose.Results:220 infants were enrolled and randomized (110 per group). 97 standard-dose and 92 high-doseinfants completed the study per-protocol. For the primary outcome, no significant difference wasobserved between groups: vaccine take occurred in 62 (67%) high-dose infants versus 69 (71%)standard-dose infants (RR 0.92, 95% CI 0.67–1.24). However, in post-hoc analysis, children with con-firmed vaccine replication had significantly increased RV-IgA responses, independent of the intervention.No significant adverse events related to study participation were detected.Conclusions:Administration of double the standard dose of an oral, live-attenuated rotavirus vaccine(Rotarix) did not improve vaccine take among infants in urban Dhaka, Bangladesh. However, improvedimmunogenicity in children with vaccine replication irrespective of initial inoculum provides further evi-dence for the need to promote in-host replication and improved gut health to improve oral vaccineresponse in low-income settings.

Development and validation of a real-time RT-PCR test for screening pepper and tomato seed lots for the presence of pospiviroids

Marleen Botermans, BioRxiv – 2020

ABSTRACT
Potato spindle tuber viroid and other pospiviroids can cause serious diseases in potato and tomato crops. Consequently, pospiviroids are regulated in several countries. Since seed transmission is considered as a pathway for the introduction and spread of pospiviroids, some countries demand for the testing of seed lots of solanaceous crops for the presence of pospiviroids. A real-time RT-PCR test, named PospiSense, was developed for testing pepper (Capsicum annuum) and tomato (Solanum lycopersicum) seeds for seven pospiviroid species known to occur naturally in these crops. The test consists of two multiplex reactions running in parallel, PospiSense 1 and PospiSense 2, that target Citrus exocortis viroid (CEVd), Columnea latent viroid (CLVd), pepper chat fruit viroid (PCFVd), potato spindle tuber viroid (PSTVd), tomato apical stunt viroid (TASVd), tomato chlorotic dwarf viroid (TCDVd) and tomato planta macho viroid (TPMVd, including the former Mexican papita viroid). Dahlia latent viroid (DLVd) is used as an internal isolation control. Validation of the test showed that for both pepper and tomato seeds the current requirements of a routine screening test are fulfilled, i.e. the ability to detect one infested seed in a sample of c.1000 seeds for each of these seven pospiviroids. Additionally, the Pospisense test performed well in an inter-laboratory comparison, which included two routine seed-testing laboratories, and as such provides a relatively easy alternative to the currently used tests.

See All The Publications

Safety Data Sheets (SDS)

Ultraplex 1-Step ToughMix, Low ROX
Ultraplex 1-Step ToughMix, ROX
Ultraplex 1-Step ToughMix

CofA (PSF)

UltraPlex 1-Step ToughMix

Description

Tough-tested, multiplex, 1-step real time PCR

Contents

Storage & Handling

Instrument Capability

Related Resources

Product Flyers

Product Manuals

Publications

Safety Data Sheets (SDS)

CofA (PSF)

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