Phusion Blood Direct PCR Kit

In order to obtain best results from Direct PCR, it is important to use a very small amount of starting materials. The recommended size is 0.3-0.5 mm in diameter. We recommend using Tissue Puncher 0.3 mm (Cat. No. F-200S), which is a convenient tool for sampling and is suitable for both plant and animal tissues.

No, extra markers cannot be added to any of the Animal Genotyping kits. All primer mixes are optimized to work in a multiplex PCR. Addition of one or more primers would require optimization of the primer concentrations and the PCR protocol.

We do not provide primers for specific locii separately. They come only in a multiplex format. If one or several loci did not amplify from the sample, faint signals can be a result of low DNA amounts or impurities. Some loci may be more sensitive towards purity of DNA used and may vary from sample to sample. On rare occasions, an animal can have a deletion in the marker region.

No, none of the genotyping kits contain an allelic ladder. Animal STR loci are highly polymorphic. Each allele has too many variants to make a universal allelic ladder.

It is very important to include a control reaction in the Direct PCR to ensure that the PCR conditions are optimal for the specific tissue or plant material used. Control primers are universal primers that amplify a small fragment from a conserved genome region. If the reaction with control primers fails, the sample material may not be suitable for Direct PCR, or PCR conditions need optimization.