However, production and purification of AAV not only represents a significant clinical manufacturing challenge ( Loo and van derWright, 2016 Halbert et al., 2018 Davidsson et al., 2020), it also limits more widespread use of this approach in the research community.īuilding on previous work, including a protocol for CRISPR/Cas9-mediated gene knock-out in human and murine T cells ( Seki and Rutz, 2018 Oh et al., 2019) and a report describing the use of linear dsDNA as repair template ( Roth et al., 2018), we set out to develop a robust, efficient, and scalable protocol for nonviral CRISPR/Cas9-mediated gene knock-in in primary human T cells. Several studies have subsequently reported high editing efficiencies using AAV-based repair templates ( Choi et al., 2019 Vakulskas et al., 2018 Dai et al., 2019). This approach was used, for instance, to insert a CAR construct into the T cell receptor α constant ( TRAC) region locus, which placed the CAR under the control of the endogenous TCR promotor, thus improving its performance ( Eyquem et al., 2017). Viral vectors, in particular adeno-associated viruses (AAV), have been used to deliver donor DNA templates for HDR-mediated target gene knock-in in T cells ( Sather et al., 2015 Wang et al., 2016 Eyquem et al., 2017 Choi et al., 2019). 4.Homology-directed repair (HDR) of double-strand DNA breaks introduced by targeted gene editing methods, such as transcription activator–like effector nucleases, zinc finger nucleases, or CRISPR/Cas9, can be used to make precise changes to a genomic sequence, including the insertion of long stretches of DNA at a defined genomic location ( Li et al., 2020 Singh et al., 2017). Questions : If you have any questions, please contact our Scientific Support Team at 0 ext. They also offer maxi-prep of plasmids, with amounts up to 30 mg. Therefore, in order to receive a price quote, work order, and ETA specific for your project, please send all your project details to Services: Additional services offered by Bio Basic include codon optimization, for no additional charge.
PUC57 SNAPGENE HOW TO
How to Receive a Price Quote: Bio Basic reviews all sequences before accepting a project. This can be supplied in any format, including WORD doc or SnapGene file.
PUC57 SNAPGENE DRIVERS
Our drivers take care of picking up the materials from your lab and shipping them to BioBasic for a fee of $40. Subcloning into one of the following vectors costs a standard subcloning price of $150:Ĭustom Vector Subcloning: Lastly, you can send your own vector to BioBasic, and they will perform the cloning for you (for the same fee of $150).
PUC57 SNAPGENE FREE
Subcloning into one of the following vectors is also free of charge:īio Basic keeps many vectors in-house, shown below. Subcloning Services: The default is blunt end cloning into pUC57 with either Amp or Kan resistance. All genes are verified by Sanger Sequencing, as well as other QC measures. What do I receive? Bio Basic supplies 4 μg of lyophilized plasmid containing your gene, and up to 10 μg for no extra charge. The proposed standard TAT gene synthesis is based on a gene with little to no complexities and with a length of about 1,000 bases. *Please note that the turnaround time is dependent on the complexity and length of a gene. Custom Genes:įrom the easiest to the most complicated gene syntheses, Bio Basic has the expertise. Our staff of experienced molecular biologists will be happy to assist you. For technical assistance contact us at call 0 ext. We will send you a price quote and, upon your approval, procure and deliver your peptide. All peptides quality confirmed by Mass Spec and HPLCįill out this form to send us the details of the custom peptide you want.Custom Peptides & Genes from Canada-based Bio-Basic Inc.
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