Bio-Rad SingleShot™ Cell Lysis RT-qPCR Kits - Manual

Introduction
The SingleShot Probes Kit prepares genomic DNA (gDNA)–free RNA directly from
cell culture in approximately 20 min for use in reverse transcription quantitative PCR
(RT-qPCR) applications. This kit is compatible with an input of 100,000–10 cells from
suspension, adherent cells, or primary cells from cell cultures. With the SingleShot
Probes Kit, gene expression analysis can be completed in approximately 2 hours from
cell culture to quantification cycle (Cq). This kit includes reagents for probe-based
RT-qPCR reactions and a probe-based qPCR control assay to optimize input cell
number and input lysate amount.
Kit Contents (100 reactions)
Cell Lysis Reagents Description
SingleShot Cell Lysis Buffer 5 ml (1 x 5 ml vial)
Proteinase K Solution 100 µl (1 x 1 ml vial)
DNase Solution 100 µl (1 x 1 ml vial)
Store all components at –20°C for up to 1 year.
Controls
SingleShot RNA Control Assay
200 reactions
SingleShot RNA Control Template Lyophilized
SingleShot Probes qPCR Control Assay HEX labeled, 200 µl
Store the control template, once resuspended, at –80°C. Store the qPCR assay at –20°C for up to 1 year.
RT-qPCR Reagents
iScript
™ Advanced cDNA Synthesis Kit for RT-qPCR 100 reactions
5x iScript Advanced Reaction Mix 1 vial
iScript Advanced Reverse Transcriptase 1 vial
Nuclease-Free Water 1.5 ml
SsoAdvanced ™ Universal Probes Supermix 5 x 1 ml vials (500 reactions)
Store the RT-qPCR products at –20°C for up to 1 year. The SsoAdvanced Supermix can be stored
at 4°C for up to 3 months.
Reagents Required but Not Provided
■ ■
Phosphate buffered saline (PBS) for washing the cells
■ ■
TE buffer (nuclease-free) pH 7.5 for resuspending the SingleShot RNA
control template
For research purposes only.
Catalog # Description
172-5090 SingleShot Probes Kit, 100 x 50 µl reactions
SingleShot
™
Probes Kit