I followed Muiltsite Gateway kit protocol to try to make 3 fragments recombination. The entry colony are good (checked sequencing). However, the LR reaction cannot correct colonies (lots colonies on plate but most of them only have 2 or 1 fragments but not all 3 together). If luck, the 3 fragment colony we got but the sequencing results are not good (bad quality and not all 3 fragments sequencing we can get). We use One Shot Mach1 (from kit) as competent cell and our destination vector is pEarlyGate 302.
Dose anyone give some suggestion. Any anyone had similar problem before and how to improvement?
Thanks.
- Badges
- Science topic
- Similar topics
- Physical Sciences
- Materials Science
- Plating
More Xin Ye's questions See All
Similar questions and discussions