Talk:20.109(S09):Induce protein and evaluate DNA (Day5): Difference between revisions
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Revision as of 15:12, 25 February 2009
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.
Group | OD WT | OD S101L | OD Chosen mutant | Pellets yellow? |
---|---|---|---|---|
Pink | 0.117 | 0.098 | 0.126 | Yes |
Yellow | 0.118 | 0.091 | 0.148 | WT and S101L bright yellow-green; Y64D paler matte yellow |
Red | 0.121 | 0.187 | 0.133 | Yes |
Orange | 0.117 | 0.087 | 0.122 | WT and S101L bright yellow-green; E31I paler matte yellow |
Purple | 0.109 | 0.075 | 0.121 | Yes |
Green | 0.106 | 0.095 | 0.072 | Yes |
Grey | 0.111 | 0.102 | 0.148 | All bright yellow-green |
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.
Group
P |
OD WT
.244 |
OD S101L
.121 |
OD Chosen mutant
.122 |
Pellets yellow?
yes |
---|---|---|---|---|
Pink | 0.105 | 0.098 | 0.105 | Yes |
Please see notes about S101L in T/R section above.