Description
Description
Use the SingleShot Cell Lysis Kit to rapidly generate cell lysates that are suitable for RT-qPCR analysis without RNA purification.
Kit works on 10 to 100,000 cells in 20 minutes while providing the following key benefits:
- Cell lysate from 10 to 100,000 cells ready in 20 minutes
- Complete removal of genomic DNA while preserving RNA integrity
- Optimal accuracy and linear dynamic range of qPCR data across genes
- Results comparable to using purified RNA, including dynamic range
- Validated with PrimePCR Assays and Panels (visit www.bio-rad.com/primepcr)
- SingleShot RNA Control included to ensure optimal cell and lysate inputs
Optimized Sample Input
The SingleShot Cell Lysis Kit maximizes cell lysate and cDNA inputs in RT-qPCR reactions:
Maximum cell lysate into RT reaction | 45% of total volume |
Maximum undiluted cDNA into qPCR reaction | 20% of total volume |
Maximum cell lysate into one-step RT-qPCR reaction | 20% of total volume |
Kit Contents
SingleShot Cell Lysis Reagents |
500 x 50 µl reactions |
Refer to the user manual for details on reaction setup and troubleshooting tips.
Cell Lines Validated for Use with the SingleShot Cell Lysis Kit
The SingleShot Cell Lysis Kit has been validated for use with a number of common human and rodent cell lines, such as all types of adherent and suspension cells, including but not limited to:
Cell Line | Property | Species |
A549 | Adherent | H. sapiens, lung carcinoma |
HeLa | Adherent | H. sapiens, cervical adenocarcinoma |
HepG2 | Adherent | H. sapiens, liver carcinoma |
NIH/3T3 | Adherent | M. musculus, embryonic fibroblast |
MCF-7 | Adherent | H. sapiens, epithelial, adenocarcinoma |
NCI-H1975 | Adherent | H. sapiens, lung adenocarcinoma |
K-562 | Suspension | H. sapiens, chronic myelogenous leukemia |
HEK 293 | Suspension | H. sapiens, kidney |
Jurkat | Suspension | H. sapiens, T lymphocyte |
PC-12 | Suspension | R. norvegicus, pheochromocytoma, adrenal |
HepRG | Primary-like | H. sapiens, hepatocytes |
RT-qPCR Step Reagents (Sold Separately)
Reverse Transcription
- iScript Advanced cDNA Synthesis Kit for RT-qPCR (172-5038)
Two-Step RT-qPCR Reagents
- SsoAdvanced Universal SYBR Green Supermix (172-5271)
- SsoAdvanced Universal Probes Supermix (172-5281)
One-Step RT-qPCR Reagents
- iTaq Universal SYBR Green One-Step Kit (172-5151)
- iTaq Universal Probes One-Step Kit (172-5141)
- Reliance One-Step Multiplex Supermix (12010220)
PrimePCR Reverse Transcription Control Assay
Use of the optional PrimePCR Reverse Transcription Control Assay in a small pilot experiment enables the determination of the optimal cell count and lysate inputs. The control can be added at the beginning to monitor the lysis step and determine the optimal cell count, or during the qPCR step to monitor RT-qPCR inhibition.
Related Products
The SingleShot Cell Lysis Kits belong to the family of SingleShot Cell Lysis RT-qPCR Kits:
- SingleShot Cell Lysis One-Step RT-qPCR Kits — including SingleShot SYBR Green One-Step Kit (172-5095) and SingleShot Probes One-Step Kit (172-5070)
- SingleShot Cell Lysis RT-qPCR Kits — including SingleShot SYBR Green Kit (172-5085) and SingleShot Probes Kit (172-5090)
- SingleShot Cell Lysis Kit — available for 100 reactions (172-5080)
Specifications
Specifications
Lysis buffer
DNase solution
Proteinase K solution
Lysis buffer