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

Introduction
The SingleShot ™ SYBR ® Green 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
™ SYBR ® Green Kit, gene expression analysis can be completed in
approximately 2 hours from cell culture to quantification cycle (Cq). This kit includes
reagents for the RT-qPCR reactions and a SYBR
® Green–based qPCR control assay
to optimize input cell number and input lysate amount.
Kit Contents (100 x 50 µl 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
200 reactions
SingleShot RNA Control Template Lyophilized
SingleShot ™ SYBR ® Green qPCR Control Assay 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 SYBR ® Green Supermix 5 x 1 ml vial (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
172-5085 SingleShot ™ SYBR ® Green Kit, 100 x 50 µl reactions
SingleShot
™
SYBR
®
Green Kit