Fragment Analysis

Fragment analysis is a capillary electrophoresis (CE) based method. CE-based assays have a simple two-step protocol (PCR followed by CE) independent of enzymatic cleavage.

CE methods have been proven to be sensitive, high-throughput and high-resolution approaches for nucleic acid analysis. It has been reported that CE methods have sensitivity and resolution that are comparable to NGS with an indel detection sensitivity of about 0.1% (Lonowski et al., 2017). Hence, they can be used in both basic research and more challenging genome editing applications such as therapeutic indel profiling.

Recently multiple CE-based assays such as IDAA, Fluorescent PCR and CRISPR-STAT have been developed in CRISPR/Cas9 genome editing studies. These are reported to be fast, sensitive, precise and cost-effective methods for mutation detection.

Similar to mismatch cleavage assay, we recommend positive control enables in your fragment analysis services for mutation detection. The positive control can be PCR fragment or PCR product with the known sequences, which typically 1 as wild type and another 1 (or more) as the target INDELs or SNPs sequences. To provide convenience to our customers, we offer gBlocks™ Gene Fragment from IDT as positive controls in our fragment analysis services for mutation detection.

Our experimental setup of fragment analysis will design according the target sequences provided by the customers. If the target sequence is not available, or customers are providing the primer design(s) to be used in the PCR service, proviso fees may be applicable depending on project requirements. Under this condition, we will offer a custom service work plan to be reviewed and accepted by the customer before confirming the inquiry.

Our regular amplification screening services before fragment analysis is now coupled with sample preparation services, allowing you to get the results in the shortest turnaround time, so that you are able to focus in the data analysis and decide the next move of your research experiments.

To maximise success for each sample preparation service, we advise our customers on how to collect and submit the samples within the acceptable terms and conditions. Such information also available in each of the respective order forms. Minimum setup fee will apply when amplification fails due to bad quality of sample that did not meet the prescribed terms and conditions.

Reference:
Lonowski, Lindsey A., Yoshiki Narimatsu, Anjum Riaz, Catherine E. Delay, Zhang Yang, Francesco Niola, Katarzyna Duda, Elke A. Ober, Henrik Clausen, Hans H. Wandall, Steen H. Hansen, Eric P. Bennett, and Morten Frödin. "Genome Editing Using FACS Enrichment of Nuclease-expressing Cells and Indel Detection by Amplicon Analysis." Nature Protocols 12, no. 3 (2017): 581-603. doi:10.1038/nprot.2016.165.
https://www.fragmentanalysis.com/tutorial

 

Specifications

Product No. Description
Genotyping Positive Control

Genotyping Positive Control

gBlocks™ Gene Fragment (251 to 500bp), Flat Price
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.

MBS-2312

PCR Amplification Screening using existing MBS ID
Include gDNA extraction, PCR reactions and Fragment Analysis.

Note: gBlocks™ is the registered trademark from Integrated DNA Technologies (IDT)