What is the sequence of the universal primer contained in the NCode™ miRNA qPCR kit?
The sequence of the universal primer is proprietary, and is not disclosed. How should I design the miRNA qRT-PCR primer to be used with the NCode™ system? Invitrogen has already designed primer sequences that you may use for each miRNA on our arrays. These can be found within the NCode™ database. The Tm of the primers were calculated using nearest-neighbor approximation based on the following reference: John SantaLucia, Jr. Hatim T. Allawi, and P. Ananda Seneviratne. Improved Nearest-Neighbor Parameters for Predicting DNA Duplex Stability. Biochemistry 1996, 35:3555-62 If you are designing a primer for a unique or novel small RNA, please consider the information below. The optimal Tm range is 58-65. For sequences with calculated Tm less than 58, either “A”s may be added to increase Tm, or modified bases could be substituted. No more than 6 “A”s should be added to any one sequence. The maximum primer length designed was 30 nucleotides. For sequences with calculated Tm greater than 65, b
