Enhance the NGS output of your pooled library screens by Cellecta´s amplicon-based NGS library prep technology

Analyzing results of pooled lentiviral CRISPR, shRNA, and barcode libraries requires targeted PCR amplification, sequencing and enumeration of the effectors from genomic DNA isolated from cells transduced with the respective library.
To streamline this process, Cellecta’s NGS Library Prep Kits provide an efficient solution for precise quantification of sgRNA, shRNA, and barcode representation in these experimental samples.

Cellecta NGS Analysis Kits contain highly specific PCR primers, optimized enzymes & reagents, and indexed amplification primers used to prepare sequence-ready libraries for multiplex NGS analysis. Primers are specific to each library/vector combination, so you simply need to choose a kit appropriate for the library you are using.

The kits come with a free data analysis software. The program demultiplexes indexed samples and aligns sgRNA, shRNA or Barcode reads from the Illumina NGS FASTQ files. The output is a spreadsheet with the counts of each effector in each sample.

NGS Prep Kits Features

Optimized protocols and reagents enable PCR reactions with a gDNA input of as high as 50 µg per reaction, which significantly reduces the number of PCR reactions required

⇒ Cost and time savings by substantially fewer PCR reactions compared to conventional low gDNA-input methods

– No background due to the reduced number of PCR cycles required

– Kits contain Index Primers for multiplexing up to 12 samples

– Data Analysis Software included: the program demultiplexes and aligns sequence data generated on the Illumina platform and scores sgRNA, shRNA- or barcode counts

– Kits are available for all Cellecta sgRNA-, shRNA-, Barcode-  and RNA-Seq Barcode Libraries as well as for GecKO and Brunello sgRNA libraries

NGS Prep Kits Workflow

– gDNA isolation from samples of genetic screens

– First Round PCR: Effector or barcode sequences are amplified using primers which are highly specific to each library/vector combination

– Second Round PCR: Introducing of indexes and P5/P7 adapters to generate sequence-ready libraries

– NGS on Illumina sequencing platform (e.g., NextSeq, NovaSeq, HiSeq, or MiSeq)

– Data analysis with provided software for demultiplexing, deconvolution, alignment, and enumeration

Full Service for Samples from Your Genetic Screen

In addition to NGS Prep Kits, we also offer full services including amplicon-based library prep, NGS on the Illumina platform, as well as data analysis for samples from screens performed with Cellecta´s sgRNA-, shRNA- and Barcode-Libraries or with GecKO and Brunello sgRNA libraries.

Simply provide cells or gDNA from your genetic screen to Nucleus Biotech, and we will deliver a spreadsheet enumerating the read counts for each of the  library elements in each sample, as well as comprehensive statistical NGS data analysis report.