The question is about S1 PFGE ; a pulsed field in which S1 is used to linearize plasmidic DNA so that the latter could migrate through pulsed elecrophoresis. i intend to run a usual PFGE, i wanted to use the same cell blocs to run the S1 PFGE, instead of the usual restriction enzyme, S1 will be placed with the blocs for 45 min (is there here any additional steps?), and the electrophoresis will be run as ususal
Is it meant to be that way ?
Thanks
Pre-incubate the PFGE agar blocks in 200 ul of 1X S1 buffer at ambient T for 30 min. Then, resuspend the blocks in 200 ul of fresh 1X S1 buffer containing 10 U of S1 enzyme, and incubate at 37°C for 10 min. Stop the reaction by suspending the blocks in 200 ul of EDTA 0.5M: incubate at ambient T for 10 min. Substitute the EDTA solution with 1X TE and leave at ambient T for at least 30 min prior to loading gel.
Thanks you, did u tried this protocol? does it work?? if it is taken from a paper would you please give me its title?? thank you for your answzr.
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