Prep of M13 RF DNA

vldnaseq at hkucc.hku.hk vldnaseq at hkucc.hku.hk
Wed Jun 22 04:02:12 EST 1994


I have an insert cloned in M13mp19 vector.  I have some difficulty in preparing
the double stranded replicative form of the clone.  The culture was grown in
2xYT until the O.D 600nm = 0.8 - 1.0 . The recombinant phage stock was added
at m.o.i = 20 . After 15 mins, chloramphenicol was added (final conc. was
15ug/ml).  The culture was harvested after 2 hr.  However, only 5 ug of ds DNA
from 20 ml culture was obtained.  Anyone know the method of producing more ds
DNA from M13 clone, please share your experience.




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