NGS Reagents for Pooled CRISPR CROP-Seq Perturb-Seq Libraries

High efficiency NGS readout of pooled CRISPR CROP-Seq/Perturb-Seq Libraries

After genetic screens perfomed with CROP-Seq / Perturb-Seq sgRNA Libraries , the frequency of each integrated sgRNA is assessed by next-generation sequencing (NGS) to determine which guides are enriched or depleted relative to the original libraries or control samples.

To streamline this process, Cellecta´s NGS Prep Kits for Pooled CRISPR CROP-Seq Perturb-Seq Libraries can be used to amplify, sequencing and accurately quantify the representation of sgRNA after running screens with Cellecta´s Pooled CRISPR CROP-Seq Perturb-Seq Libraries

The Kits contain all reagents necessary reagents, including highly specific PCR primers for efficient amplification of sgRNA, and sgRNA-barcode cassettes as well as indexed primers to prepare sequence-ready libararies for multiplex NGS analysis.

NGS Library Prep Kit 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

– Free Data Analysis Software included: the program demultiplexes, aligns, and scores Illumina platform sequence data

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

Workflow:

– gDNA isolation from samples of genetic screens

– First Round PCR: sgRNA sequences are amplified with primers  that are highly specific to the CROp-Seq library vector backbone (pScribe).

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

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

– Data analysis with included software: the program demultiplexes indexed samples and aligns sgRNA reads from the Illumina NGS FASTQ files. The output is a spreadsheet with the counts of each sgRNA in each sample.

In addition to NGS Prep Kits, we also offer Cellecta’s next-generation sequencing and analysis services for Cellectas sgRNA-, shRNA-, Barcode-  and RNA-Seq Barcode Libraries as well as for GecKO and Brunello sgRNA libraries.

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

NGS Reagents for CROP-Seq / Perturb-Seq sgRNA Libraries are tools to amplify and sequence sgRNAs, after running screens with pooled lentiviral libraries. For CRISPR screens, the frequency of each integrated sgRNA is assessed by next-generation sequencing (NGS) to determined which ones are enriched or depleted relative to the original libraries or control samples.
Nucleus Biotech provides NGS Prep Kits for various libraries to meet your needs. Each contains the appropriate primers for particular library/vector combinations to generate sequencing-ready samples. Simply select the Prep Kit for your screen and order.