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IF you calculate∆Ct = Ct[Housekeeping]-Ct[Target] ... and ∆∆Ct = (∆Exp.)-(∆Control)THEN∆∆Ct is a log-fold-change.IF you calculate∆Ct = Ct[Target]-Ct[Housekeeping] ... and ∆∆Ct = (∆Exp.)-(∆Control)THEN-∆∆Ct is a log-fold-change(logs to the base 2).IF ...

Measuring optical density (OD) using a spectrophotometer is often problematic if the physics behind the method is not understood. For example, the relationship is between biomass concentration and OD, not cell number and OD. This is because larger ce...

It is very easy. For instance, you want to make a 5% solution in TBS-Tween. You just put 2.5 g BSA in 50 ml of TBS-Tween, you put the sample at 4 C and after 10 minutes it is dissolved. No stirring, no mixing.It is worthing noting that you should not...

PyMOL sometimes displays bonds which are not correct and can be confusing. To remove a bond between to atoms, first set the selection argument to "Atoms", then select the first atom involved in the bond, right click and goto "actions&q...

If you want to relate expressions among several groups, it's best to show dCt. If the comparisons at all done to the same reference, best show ddCt.Never ever use simple barcharts (as used so terribly often in biomedical papers). If you have smal...

1. The technical replicates.A technical replicate is when you test the same sample multiple times - it's used to test the variability in the testing protocol itself.The reason you do technical replicates is to make sure they are almost identical....

There are many factors that may affect the protein binding:0. The His tag is incorporated into the tertiary structure of the protein and inaccessible to the Ni column. Try a batch under denaturing conditions (8M Urea or 6 M Guanidine) to check this. ...

It is much better to use g (the capital G is not the correct unit) as a unit for centrifugation steps, which refers to the acceleration applied to your samples (so 10,000 g means 10,000 times Earth's gravitational force). Also, 'g' is the...

To get good PCR yield and reduce non-specific products, how much DNA template is generally used in a 50ul PCR reaction?Generally, for low complexity templates (i.e. plasmid, lambda, BAC DNA), use 1 pg–10 ng of DNA per 50 µl reaction.For higher comple...

The point of the glycerol used for storage of bacteria is preventing freezing damage, such as ice crystal formation. At lower glycerol concentrations like 10-20% the frozen stock will be fairly solid at -80C, while at higher concentrations (30-50%) i...