Intended Use

• The Master Mix is used for real-time qualitative and quantitative RT-PCR amplifications with SYBR Green dye for extraction-free RNA samples.

• The master mix is a premixed, 2X concentrated solution that has all the components except for gene-specific primers and RNA template.

Kit Characteristics

• The master mix contains extra more redundant amounts of the RTase and DNA polymerase, specifically designed to overcome endogenous inhibition in extraction-free RNA samples.

• For the reverse transcription step, this kit uses a highly efficient Thermophilic Reverse Transcriptase (US patent pending), which is a thermophilic type A polymerase, with optimal temperatures of 60-62°C.

• The RTase is easily heat-inactivated at ≥90°C for 1min.

• The RTase efficiently synthesizes a complementary DNA strand on RNA template from a gene-specific primer, ≤1 unit per 20μL of reaction.

• The RTase reversely-transcribes single digit copies of target RNA molecules consistently.

• The kit also contains Taq-Fast DNA polymerase which extends more than 300 bases with short cycling program.

• The concentrations of the primers are variable depending on assay designs and thermocycling protocols (Table 1).

• The preferable PCR product size is ≤150bp.

• The kit has three formulations of No ROX, Low ROX or High ROX concentrations for your choice.

User Manual

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