AccuStart II PCR ToughMix is a 2X concentrated ready-to-use reaction cocktail for PCR amplification of DNA templates that overcomes many known inhibitors of PCR often present in crude samples extracted from environmental specimens, plant tissues, or animal tissues. The only user supplied components are DNA template and molecular grade water. A key component to AccuStart PCR ToughMix is an ultra pure, highly processive thermostable DNA polymerase that is combined with high avidity monoclonal antibodies. These antibodies bind the polymerase and keep it inactive prior to the initial PCR denaturation step. This enables specific and efficient primer extension with the convenience of room temperature reaction assembly. Similar to Taq DNA polymerase, the activated polymerase in AccuStart II PCR ToughMix possesses 5’>3′ DNA polymerase activity and a double-strand specific 5’>3′ exonuclease. The polymerase does not have 3′-exonuclease activity and is free of any contaminating endo or exonuclease activities. PCR products generally contain non-templated dA additions and can be cloned using vectors that have a single 3′-overhanging thymine residue on each end.   GelTrack Loading Dye is a mixture of blue and yellow electrophoresis-tracking dyes that migrate at approximately 4kb and 50 bp. This optional component simplifies post-PCR analysis with gel electrophoresis and eliminates potential for cross contamination by enabling direct transfer of PCR products to the gel sample wells.