understanding Trackit X174 RF DNA/HaeIII Fragments - confused (Apr/05/2005 )
I am going to use this product to quantify mtDNA but do not understand how to use it. I understand that based on intensity I can quantify the DNA present but I do not understand how to find out how many ng of DNA are represented by each band. If I add up the base pairs for each band I get 5386. Then I divide the number of bp of each band by the total # (5386). Now I don't know what to do. And how do I take the volume of the ladder used into consideration. Thanks in advance.
normally, different amounts of ng are used for ezach band in order to have a good ladder with all bands vis
TrackIt™ X174RF DNA/Hae III Fragments are supplied at 0.1 µg/µl in 10 mM Tris-HCl (pH 7.5), 10 mM EDTA (pH 8.0), 0.06% XCFF, 0.6% tartrazine, and 5% glycerol.
that means that each fragment has a concentration of 0.1 µg/µl. Hence, if i assume ou use 2µl of track it X174 RF DNA/Hae III ladder, you'll get 200ng for each band.