Reverse Transcription Kit is intended for reverse transcription using enzyme MMLV reverse transcriptase. This enzyme converts RNA to cDNA (complementary DNA) in one step (without denaturation), you only have to add your template RNA.
Parameters of the product
- Master Mix is 2x concentrated.
- One vial contains 1 ml of Master Mix (this amount is sufficient for 100 reactions of 20 µl). PCR Master Mix contains hot-start Taq DNA polymerase, reaction buffer, dNTP, MgCl2, fluorescent dye (with detection in FAM/Sybr channel) and other components needed for PCR amplification with subsequent HRM analysis.
- PCR Master Mix does not contain any passive reference dye.
- If the real-time PCR cycler requires normalization with a passive reference dye, it can be added to the reaction mixture separately (see Passive Reference Dye, Cat. No. 3010 – on request)
|Initial denaturation||95 °C||3 min|
|Denaturation||95 °C||5 s||30-50 cycles|
|Annealing+elongation (+fluorescence acquiring)||55-65 °C||10 s||30-50 cycles|
|Extension||72 °C||20 s||30-50 cycles|
|Denaturation||95 °C||2 min|
|HRM analysis/Melt Curve (+fluorescence acquiring)||50 °C||20 s|
|HRM analysis/Melt Curve (+fluorescence acquiring)||70-95 °C||5s||rise by 0,1-0,2 °C|
|gb PCR Master Mix by application
|end-point PCR, common PCR amplification|
|real-time PCR without probes||
|real-time PCR with hydrolysis probes|
|real-time PCR with LNA probes|
|real-time PCR with hybridization probes|
|real-time PCR with High Resolution Melting Analysis||
|real-time PCR with low DNA samples|
|real-time PCR with inhibited sampels||