The genetic and molecular biology teams have developed a number of novel recombinant constructs. There is a $100 cost recovery fee for up to 5 constructs per order, and an Institutional Materials Transfer Agreement (MTA). We ask for an acknowledgment in any publication that uses this resource. The receiving lab is to cover the cost of mailing by supplying a FEDEX account number.
Constructs are supplied as 200ng of desiccated DNA for transformation. They have been sequenced and to the best of our knowledge are good. However, use of this material is at the investigator’s own risk. We recommend all constructs are sequenced, prior to use. Any feedback would be welcomed. We hope that this resource will be helpful and will accelerate the pace of research to develop new treatments for neurodegenerative disease. We would be pleased to collaborate and share our experience in working with these tools. However, there are no restrictions or obligations on use.
Maps for some plasmids may be available upon request (SAM, BAM, Geneious or Fasta extensions, if available sequence may be provided as a text file)
At least 200ng of DNA will be sent dried on Whatman paper. Protocol to remove DNA from paper is as follows:
- Cut around outlined circle and place in micro centrifuge tube
- Add enough TE or ddH2O to cover the paper (20-50uL depending on size of circle)
- Incubate at room temperature or 52 degrees for 10 minutes (52 degrees may produce a higher yield for larger plasmids – greater than 5kb)
- Spin down at high speed for 30 seconds to elute
- Pipette out any liquid (this will contain your DNA) and transform 10-15ul in competent bacteria. Save any extra liquid in case transformation doesn’t work.
- Plate 100 ul of transformation. Spin down rest of transformation, remove supernatant. Resuspend bacteria pellet in 100ul of medium. Plate entire bacteria pellet on another plate.