Solutions for the whole workflow from Laser microdissection to RNAseq
1. Sample prep with Laser Microdissection
2. The importance of accurate cell counting – NEW!
3. Sample stabilization with CellCover – NEW!
4. Whole genome amplification
5. cDNA synthesis
6. Single-cell Methylation Sequencing
7. RNAseq solutions – NEW!
1. Pure starting material for downstream analyses with Leica Laser Microdissection!
Laser Microdissection (LMD, also known as Laser Capture Microdissection or LCM) enables users to isolate specific single cells or entire areas of tissue. Powered by a unique laser design and dynamic software, Leica LMD systems allow users to easily isolate Regions of Interest (ROI) from entire areas of tissue down to single cells or even subcellular structures such as chromosomes.
LMD is typically used in genomics (DNA), transcriptomics (mRNA, miRNA), proteomics, metabolomics, and next generation sequencing (NGS). Researchers in neurology, cancer research, plant analysis, forensics or climate research rely on this method. Furthermore, LMD is a perfect tool for live cell culture (LCC), for cloning and re-cultivation, manipulation or downstream analysis.
Gravity for Cleanliness
Your downstream analysis relies on contaminate-free isolates. This is why Leica LMD systems collect dissectates via gravity. Their unique laser-guidance based dissection method preserves the integrity of your isolates – contact-free and contamination-free.
2. Accurate cell counting with Cellometer K2 Automated Cell Counter
Accurate cell counts are critical in sample preparation for single-cell sequencing. AO/PI fluorescent viability staining and automatic de-clumping can minimize counting errors due to debris, dead cells, cell clustering etc.
Cellometer K2 Automated Cell Counter can be customized to handle a variety of cell types including: low or high concentrations of primary cells, insect cells, cell lines, fragile cells, clumpy cells, samples with large amounts of debris, and more.
- Requires only 2-20 ul input material
- Takes only 30 seconds per count
- Read the Single Cell Application note to learn more!
3. Stabilize your cell & tissue samples with CellCover
Anacyte’s CellCover is the only reagent that allows parallel storage of proteins, RNA, and DNA in their cellular context, maintaining cellular shape integrity without chemical crosslinking.
- Stabilize RNA, DNA and proteins simultaneously for several days
- Straight forward RNA isolation
- No freezing necessary
4. For whole genome amplification WGA of your limited samples: Lucigen NxGEN Phi29 DNA polymerase
- Complete amplification of limited samples before PCR or NGS
- Excellent strand displacement activity, fidelity and speed
- Great performance with 70% lower price
5. Do more with less – Produce cDNA from precious total RNA or cells: Lucigen MessageBooster cDNA synthesis kits
- Fast: Amplify RNA and produce cDNA in only 1 day
- Unbiased Amplification: Preserves the relative transcript abundance of a sample
- Use samples you couldn’t before, produce enough cDNA for multiple assays
- Detect even low-abundance transcripts in RNA from a single cell or 10 pg of total RNA
6. For Single-cell Methylation Sequencing: Swift Biosciences Accel-NGS Adaptase Module
- Constructs NGS libraries from bisulfite-converted, single-stranded DNA from single cells
- Optimized to maximize the recovery of DNA
- Three-dimensional indexing for high-throughput sequencing 384-plex
- Described in Science paper
7. RNAseq solutions for single-cells
a) Expression profiling of single cells: Lexogen LUTHOR 3’ mRNA-Seq Library Prep Kit
Unprecedentedly sensitive 3’mRNA seq lib prep solution, which enables the expression profiling from very low input amounts.
Input 1-100 cells (from e.g. FACS or suspensions), or 10pg total RNA. Detect and sequence twice as many genes per cell, cost-effectively on the Illumina platforms.
b) For whole transcriptome analysis of single cells: NuGEN Ovation Solo
- A novel & unique end-to-end solution enables whole transcriptome analysis of ultra low input levels (1-500 cells/10pg-10ng total RNA input)
- With targeted depletion of rRNA and customizable for other unwanted transcripts
- Species-specific depletion done on library level; saving your sample
- AnyDeplete technology increases the dynamic range, reduces sequencing costs and simplifies data analysis -> More data with less sequencing!
- Designs already available for Human, Mouse, Drosophila