Preserving the integrity of inverted terminal repeat (ITR) sequences in adeno-associated virus (AAV) plasmids is important for the production of recombinant AAV (rAAV) vectors. Azenta Life Sciences, formerly GENEWIZ’s proprietary AAV plasmid preparation service delivers superior quality for midi to giga-scale AAV vectors, while maintaining the integrity of ITR regions. Our new process includes pre- and post-preparation quality controlled (QC) steps that leverage Azenta's AAV-ITR Sanger sequencing capabilities for full sequence verification and ensures delivery of intact ITR clones for your research.
Truncated ITRs can reduce the yield of full viral capsid production and increase generation of undesirable non-rAAV encapsidated DNA. Unfortunately, AAV plasmid ITRs are frequently mutated during plasmid production in bacterial cells. Azenta’s AAV plasmid preparation service incorporates a two-step QC process, including pre-prep and post-prep QC. The pre-prep QC step assesses your sample configuration for the presence of full-length intact or truncated ITRs, or a mixture of both. This allows selected propagation of intact ITR sub-populations only, while the post-prep QC step ensures integrity of ITR regions in the final plasmids delivered to you. For whole plasmid sequencing, utilize our primer walking service with AAV plasmid preparation.
*Plasmid DNA is currently the only starting material accepted
Note: Screening and selection of intact ITR clones incurs additional cost
**6-day turnaround time for samples that have previously been ITR sequence verified at Azenta.
Azenta has recently developed a proprietary AAV plasmid preparation process, which significantly improves the chance of maintaining ITR integrity, even when other commercially available competent cells growing in low temperature have failed. This process can also isolate clones with full-length ITR from a sample mixture containing intact and truncated ITRs.
Azenta has developed a high-quality, direct Sanger sequencing method that reads through both intact and commonly mutated inverted terminal repeat (ITR) regions of adeno-associated virus (AAV). This method is an effective tool for assessing the integrity of ITRs in AAV plasmids.
The adeno-associated virus (AAV) is a powerful vehicle for gene therapy; however, working with AAV vectors can be challenging. Spontaneous mutations in the inverted terminal repeat (ITR) regions can accumulate during plasmid propagation in bacteria. Find out how our scientists have optimized the AAV plasmid preparation process to deliver 100% accurate constructs.
Working with AAV vectors can be challenging. In this on-demand workshop & roundtable discussion, you’ll gain a better understanding of the AAV development pipeline for gene therapy research and learn how to optimize upstream and downstream processes including AAV synthesis, sequencing, bioinformatics, and storage.