CelluLyser™ Micro Lysis and cDNA Synthesis Kit

Fast and easy 1-tube cell lysis and reverse transcription of 1-500 cells
The CelluLyser™ Micro lysis buffer has been de novo designed to provide improved mRNA retrieval even for very low expressed genes, enhanced stabilization of mRNA, minimal degree of dilution and high compatibility with downstream reagents.

By generating cDNA directly from cell lysate without any washing steps higher yields are achieved, and to avoid losses of material the entire procedure can be performed in a single tube (or 96/384-well plates) without any sample transfer.

The CelluLyser™ Micro lysis reagent has been developed for fast and simple cDNA preparation for single cell applications including very small samples (FACS sorted single cells, cell cultures, dissociated tissue).

Description
The CelluLyser™ Micro's properties make direct lysis and analysis of small cell numbers more accurate, flexible and feasible.

The CelluLyser™ Micro buffer and cDNA synthesis kit has been developed for fast and simple cDNA preparation for single cell applications including very small samples: FACS sorted single cells, cell cultures, dissociated tissue.

By generating cDNA directly from cell lysate without any washing steps, higher yields are achieved, no bias in transcriptome is introduced and considerable amount of time is saved compared to standard column based purification (Svec et al. 2013). To avoid losses of material the entire procedure can be performed in a single tube (or 96/384-well plates) without any sample transfer, just adding reagents.

The whole process can be easily automated for high-throughput applications.

Features

• Lysis buffer optimized for direct cell lysis from a single cell up to 500 cells in only 5 µl volume
• Optimized to preserve and stabilize released RNA
• Compatible with downstream reagents for reverse transcription
• Compatible with TATAA Universal RNA and DNA spikes for quality control of the workflow

Cell types
The CelluLyser™ Micro buffer has been developed to be a mild lysis solution. It has been verified on many types of in vitro and in vivo viable cells including astrocytes, microglia, fibroblasts, lymphocytes (B-cells, T-cells), leukemia cell lines (K562, HEK293), monocytes, breast cancer cells, sarcoma cell lines and embryonic stem cells.

Contents

• CelluLyserTM Micro buffer: 200 rxn à 5 µl, 1.0 ml (1X)
• GrandScript cDNA synthesis kit 200 rxn à 10 µl RT
• 400 µl GrandScript RT Reaction Mix (5X)
• 100 µl GrandScript RT enzyme
• Nuclease free water