Next Generation Sequencing (NGS)
NGS enables indel detection in both mixed populations and clonal cell lines. Its ability to process a large number of samples (samples are pooled together for a high-throughput sequencing run) and simultaneously screen off-target changes makes it a popular choice among researchers. In addition, as in all sequencing approaches, the nature and diversity of mutations are revealed
The data obtained from targeted amplicon sequencing is highly sensitive with detection levels as low as 0.01% (Hendel et al. 2015). This means that the researcher can be relatively certain that their samples do not contain off-target mutations if they are undetectable using this technique.
Besides being able to detect if all alleles of a gene were correctly edited, information on indel location and whether a cell population is truly monoclonal can be derived from NGS data.
Hendel, Ayal, et al. “Quantifying on-and off-target genome editing.” Trends in Biotechnology 33:2 (2015)
|MBS-2201||Experimental Setup: Genotyping/ Mutation & Variant Screening using qPCR.
Charge per mutation or variant, excludes cost of probe/ SNP assay/ assay positive control.
The cost of probe/ gBlock™/ SNP assay kit to be charged separately.
PCR Amplification Screening using existing MBS ID
Deep Sequencing for PCR amplicons
- Sample QC
- Library preparation and QC
- Illumina MiSeq 250 PE/ 300 PE run
- Data Output: 200,000 reads per sample
- Standard Bioinformatics Analysis (NGS-0606)
Standard Bioinformatics Analysis included: -
- Data QC and trimming
- Variant Calling
Library Preparation - Illumina platform
- Sample QC and Report
- Illumina Truseq PCR Free/NEB DNA Library preparation
- Library QC
Library Pooling Service
- Price per pool
Illumina Sequencing (Per Lane)
- Options of read length (choose one)
150 PE/ 250 PE/ 300 PE run
- Options of data output per lane (choose one)
(i) MiSeq 150 PE Nano v2: 300Mb
(ii) MiSeq 250 PE Nano v2: 500Mb
(iii) MiSeq 300 PE v3: 12Gb
(iv) HiSeq 150 PE: 110Gb
(v) NovaSeq 150 PE: 850Gb
Note: gBlocks™ is the registered trademark from Integrated DNA Technologies (IDT)