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How to interpret result of fold change in qPCR?

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 ...

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How to exactly measure the OD600 of E.coli suspension?

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...

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How to dissolve BSA powder?

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...

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How to publish qPCR data in a bar graph?

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...

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Why do His tagged protein not binding to Ni beads?

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. ...

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How to convert centrifuge RPM to RCF or g force?

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...

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