◎ Reverse transcription and quantitative PCR can be completed in one step in one reaction tube, and the operation is simple and fast;
◎ Using chemically modified hot-start polymerase, the reaction specificity is high, and nonspecific amplification is effectively avoided;
◎ The reaction buffer is fully optimized, with high sensitivity, and the target fragment in 1pg total RNA can be detected at the lowest level, which is especially suitable for the detection of low-copy RNA.
◎ The output of RT-PCR can be obviously improved by adding patented live elements of EMI. The CT value is generally 3-5 cycles earlier, and the S-shaped curve is typical.
Description: 1, BIOGHSC? Super? The probe mixture contains dNTP mixture and Mg2+, BIOGHSC? Super? The enzyme mixture contains BIOG super reverse transcriptase, BIOG hot-start polymerase and fluorescein. 2. Generally speaking, the final concentration of primers in the reaction system is 0.3μM, which can get better amplification effect. When the reaction result is unsatisfactory, the primer concentration can be adjusted in the range of 0. 1- 1.0μ m ... 3. The final concentration of 3.TaqMan probe can be adjusted between 0. 1 micron and 0.45 micron. 4. QCPR has high sensitivity, and the accuracy of template amount has great influence on the results. It is suggested to add the template after moderate dilution, or use 50? L reaction system. 5. The template volume should not exceed 10% of the reaction volume. 6. Please select the length of the amplification product within the range of 100bp-500bp, preferably 100-200bp. ? 7. When preparing the reaction system, avoid violent stirring to prevent bubbles. 8. reaction conditions: reverse transcription reaction15min at 42℃, heating at 95℃ for 5min to activate hot-start DNA polymerase, and then heating at 95℃? 15 second, 60℃? 35 seconds, 35-40 cycles. The reaction conditions can be appropriately adjusted according to the size of the target fragment and the Tm value of the primer. 8. If the template has a complex secondary structure, the reaction temperature can be increased to 55℃, which is helpful to improve the yield.