Q: Are these kits compatible with DNA from paraffin embedded tissues?
A: These kits can be used on any DNA sample, including DNA that has been extracted from paraffin embedded tissues.
Q: Can sheared DNA be used with these assays?
A: The standards are genomic DNA but the DNA size won’t matter with this assay as long as the DNA is denatured to single strands. The DNA coated plates will bind the same amount of DNA regardless of size.
Q: Can plasmid DNA be used with these kits?
A: Plasmid DNA can be used with both of these kits, since the structure of the target marker will be the same regardless if it is from plasmid or genomic DNA.
Q: Is this kit suitable for use with bacterial cells?
A: Since this kit uses isolated DNA samples, it is suitable for use with DNA extracted from bacteria.
Q: Can you recommend a DNA extraction kit?
A: Any DNA extraction method or commercial kit is compatible with our UV induced DNA damage kits. At the end of the DNA extraction kit protocol, the DNA pellet can be resuspended in either the kit buffer (Tris-HCl) or water because the DNA will be diluted to 2 µg/ml in PBS as part of the assay protocol.
Q: What is the UV source used to induce damage?
A: We use a Sylvania 30 watt germicidal (G30T8) bulb for UV light treatment.
Q: How do I make the standard curve and calculate my results?
A: The best way to determine concentrations from a standard curve is to use a 4-parameter curve fitting program, but if you don’t have this software it can be done using Excel. Graph the standard curve and generate a linear trendline. The standard curves generated with ELISAs are not typically linear, but a linear curve can be created by eliminating the upper and lower values of the curve, as long as the sample values fall within this range. The middle part of the standard curve is the most sensitive and is the best part to use for quantifying samples. The equation of the trendline can be used to calculate the concentrations of unknowns by solving for x in y=mx+b, which is (OD value-b)/m for linear lines.