Table 2. Results of thermal denaturation experiments in different types of buffers for different types of oligoribonucleotides.
Medium salt buffer |
Medium salt buffer with FA |
Low salt buffer with FA |
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Probes analyzed | Sequence | RNA complement Tm (°C) | RNA complement Tm (°C) | RNA complement Tm (°C) | ||
Ref | 5’-CTGGAGAGACTAAGCCCTCCAA-3’ | 64 | 69 | 68 | ||
HP_LNA_PO | 5’-FAM GALCTLAALGCLCCL -3’ | 78 | 69 | 67 | ||
HP_LNA_PS | 5’-FAM G*AL *C*TL *A*AL *G*CL *C*CL -3’ | 77 | 68 | 66 | ||
HP_LNA/2OMe_PO | 5'- FAM GLACTLAAGLCCCL-3’ | 79 | 66 | 67 | ||
HP_ LNA/2OMe _PS | 5'- FAM GL* A*C*TL*A*A*GL*C*C*CL*-3’ | 78 | 63 | 66 |
The DNA oligonucleotide probe reference (Ref) has the following sequence: 5’-CTGGAGAGACTAAGCCCTCCAA-3’. The RNA complementary oligonucleotide has the following sequence: 5’-UUGGAGGGCUUAGUCUCUCCAG-3’. LNA nucleotide monomers are represented with L superscript, 2’-OMe-RNA monomers in boldface letters, DNA nucleotides in capital letters, and phosphorothioate linkages by the symbol*, FAM- Fluorescein, FA- Formamide.