SingleShot Cell Lysis Kit, 100 x 50 µl rxns #1725080

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Overview

Pkg of 1, kit includes 5 ml (1 x 5 ml) of cell lysis buffer, 100 µl of proteinase K solution, and 100 µl of DNase solution, for 100 x 50 µl reactions.

Safety Data Sheet

Safety Data Sheet

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
SingleShot Cell Lysis Buffer 
SingleShot Proteinase K Solution 
SingleShot DNase Solution

100 x 50 µl reactions
5 ml (1 x 5 ml vial)
100 µl (1 x 1 ml vial)
100 µl (1 x 1 ml vial)

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 RT-qPCR 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

Storage at –20°C
12 months:
Lysis buffer
DNase solution
Proteinase K solution
Storage at 4°C
6 months:
Lysis buffer