Traditionally, Southern blot analysis is the method of choice to satisfy FDA’s requirements of characterizing cell line genetic stability. Avance Biosciences is now offering two NGS approaches to analyze the transgene insertion sites.
The first approach performs whole genome sequencing on gDNA extracted from the cell line of interest. Illumina’s PCR-free method is used for library preparation with the sequencing performed on a HiSeq. The paired-end sequencing reads generated are analyzed using a proprietary NGS analysis pipeline involving multiple rounds of mapping and assembly to elucidate the insertion junctions. The main advantages of this approach are that a reference genome sequence is not required and this method can be easily validated.
The second approach involves ligation of an adaptor followed by a nested PCR to pull down sequence fragments with insertion junctions. Sequencing can be performed on an Illumina’s MiSeq, and the paired-end sequencing reads analyzed to identify insertion junctions. The main advantage is that many samples can be analyzed easily once the assay is established. This method does require knowledge of the host genome sequence.
Our Customers Say…
From the preliminary conclusion of this work, we are already pleased to thank you for your customer-oriented approach and proactive communication in the course of this study. We are happy to see the profound difference in business mindset between your organization and our previous vendor…
Project Manager, Belgium
Thank you so much for successfully completing this important project for us on time. We are very satisfied with the final report and thank you for taking the extra effort to customize the report format. We will definitely use your lab for future projects.
Program Manager, California
It has been a pleasure working with you and your team. The project was carried out to the highest standard and delivered in a timely manner. I will definitely recommend Avance Biosciences’ services to colleagues and friends.
Senior Scientist, Singapore