Anointed as Nature’s 2013 Method of the Year, single cell sequencing allows you to interrogate genomic features in single cells versus the heterogeneity found in entire cellular populations. Genomic features such as single nucleotide and copy number variations (SNVs, CNVs) can be lost with population based averaging. By sequencing individual cells, variability in even homogenous populations of cells can be examined. Techniques to prepare single cells for sequencing typically rely on whole genome amplification (WGA), template switching, and multiple annealing PCR primer amplification strategies. More details about these techniques, can be found in our blog post: Sequencing Genomic Content from a Single Cell.

Genohub has experience designing single cell DNA experiments. Contact us to take advantage of our complementary consultation service. You can also search for and identify providers who offer single cell DNA-Seq services.

See Genohub's up-to-date list of available Single cell DNA-seq library prep services.

Single Cell DNA-Seq Kits

Qiagen REPLI-g Single Cell Kit

The REPLI-g kit utilizes multiple displacement amplification (MDA) to amplify DNA isolated from single cells. Kit claims include accurate, unbiased profiles of genomic loci and high library yields (40ug/uL).

Protocol Overview:

1. Start with single cells or purified gDNA
2. Alkaline denaturation
3. Buffer neutralization
4. MDA amplification

Rubicon Genomics PicoPlex DNA-Seq Kit

The PicoPlex DNA-seq Kit is designed to prepare single cell DNA libraries that are compatible with Illumina platforms. The entire library preparation workflow happens in a single tube. Cells are lysed, quasi-random primers pre-amplify the DNA selectively, and a final PCR amplification adds Illumina compatible barcodes. Users can start with as little as 1-10 cells or 6 pg – 60 pg of isolated DNA.

Protocol Overview:

1. Start with template DNA
2. Anneal and extend using random non-complementary primers
3. Denature, re-prime, extend
4. Denature, recycling template
5. Amplify