Method for obtaining information on formation of double-stranded nucleic acid
A technology of double-stranded nucleic acid and nucleic acid chain, which is applied in the field of obtaining double-stranded nucleic acid formation information, can solve problems such as high cost, and achieve the effect of reducing background signal and high detection sensitivity
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Embodiment 1
[0101] To the solution containing the nucleic acid strand N1 of the 18mer base sequence fluorescently labeled with fluorescent dye "LightCyclerRed 640" (product of Roche Diagnostics K.K.), the solution comprising the nucleic acid strand N2 having a base sequence complementary to the nucleic acid strand N1 was added and "SYBR TM Green I", followed by mixing. After heating the resulting mixture at 95°C for 1 min and then incubating at 20°C for 5 min, the TM At 498nm of the excitation wavelength of Green I", the fluorescence spectrum was measured using a fluorescence spectrometer. During the measurement, the interaction solution (reaction solution, interactionsolution) was controlled to be kept at 20°C.
[0102] The experimental conditions are shown in Table 1.
[0103] Table 1
[0104] Solution composition
Embodiment 2
[0113] A solution containing a nucleic acid strand N3 (base sequence: TGCCCACTATTAAGGAAGG) that does not form a double-stranded nucleic acid with the nucleic acid strand N1 and "SYBR TM Green I", and the fluorescence spectrum was measured. Experimental conditions such as solution composition were set similarly to those in Example 1.
[0114] The fluorescence spectrum of embodiment 2 is shown in Figure 7 middle.
[0115] It can be understood that although no double-stranded nucleic acid is formed, the recognition of "SYBR TM The fluorescence of the emission wavelength (520nm) of GreenI ", and observe background signal. Because background signal reaches maximum at 520nm, then gradually reduces with wavelength shift, therefore thinks that in the method according to the present invention, detects the fluorescence of target fluorescent dye. Select to reduce background signal.
[0116] The comparison of the fluorescence intensity is then performed by changing the kind of fluor...
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