Talk:20.109(S09):Induce protein and evaluate DNA (Day5)





Improved Fsp Digest


 Lane 1 is the 1 Kbp ladder, lane 5 the 100 bp ladder.

Lanes 2, 3, and 4 are uncut IPC (WT), digested IPC, and digested IPC-S101L, respectively.

Samples were digested overnight, as FspI appears to digest somewhat weakly otherwise. 

TR O/N cultures and Gels
The table below lists the OD600 values of 1:10 dilutions of the cultures grown overnight, as well as the pellet observations.



Note: 1% gels were run for 45 min, 1.2% gels for 60 min, both at 100V.



 There seems to have been a contamination or other strange issue with the S101L DNA and/or FspI enzyme. For your final reports, please use my old data, which is posted above. Unfortunately, some of the bands are faint (this experiment pre-dates our modern gel doc system). Lane 1 is parent IPC (only one form of the plasmid is visible), lane 2 is the ladder, lane 4 is the parent digested with FspI, and lane 8 is the S101L mutant digested with FspI. The short band is very faint, but it is present. I will have the original photo available for viewing for anyone who wants to clearly see the short band. At right is a cropped version for easier viewing. '''Update: Upon reflection, most of the S101L data does not look nearly as bad as I thought. As you interpret your data, keep in mind what are all the possible band sizes you could get. What if the enzyme only partially digests? Nevertheless, I will try to re-run this experiment to get a better photo for your reports in the coming days.'''



WF O/N cultures and Gels
The table below lists the OD600 values of 1:10 dilutions of the cultures grown overnight, as well as the pellet observations.









Please see notes about S101L in T/R section above.