This is a protocol Allan DeBono used when cleaning BigDye 3.1 sequencing reactions for sequencing at NAPS. In both of our hands it worked consistently to confirm vector inserts. An advantage for this protocol is that it takes 5-10 minutes to preform depending on how many reactions are being run (and how fast of a pipetter you are). It also utilizes spin columns that can be re-used after autoclaving. Continue reading
The following protocol is for producing electro-competent cells of agro-bacterium. Currently A. tumefaciens agropine-type strain EHA-105 (Hood et al.,1995) is used for transformation of sunflower and arabidopsis. Harvesting new competent cells takes 4 days to complete with the 4th day being the most labour intensive. It’s advised to begin early in the morning on the 4th day as the length of culture, purifying, and aliquoting can take several hours.