«ALSENSE FLU» Reverse-Transcription PCR Kit

The prin­ciple of op­era­tion of the Re­verse-Tran­scrip­tion PCR Kit «AL­SENSE FLU» is to con­duct re­verse tran­scrip­tion and real-time poly­merase chain re­ac­tion (qP­CR) with hy­brid­iz­a­tion­fluor­es­cence de­tec­tion in one tube. The re­verse tran­scrip­tion re­ac­tion is per­formed to form com­ple­ment­ary DNA (cDNA) on the tar­get RNA mat­rix. In the PCR step, the res­ult­ing cDNA is amp­li­fied mul­tiple times. De­tec­tion of amp­li­fic­a­tion products is per­formed dur­ing the ana­lys­is. For this pur­pose, spe­ci­al fluor­es­cent tags (probes) are ad­ded to the re­ac­tion mix­ture, along with primers and oth­er re­ac­tion com­pon­ents. Each DNA probe car­ries a fluor­es­cent tag and a fluor­es­cence quench­er. When a spe­cif­ic product is formed, the DNA probe is des­troyed and the quench­ing agent stops act­ing on the fluor­es­cent tag, which leads to an in­crease in the fluor­es­cence level. The hy­brid­iz­a­tion probe con­tains both a fluor­es­cent dye and a fluor­es­cence quench­er in its struc­ture. In the case of a pos­it­ive re­ac­tion, the hy­brid­iz­a­tion probe is an­nealed on cop­ies of a giv­en DNA site and in the next cycle of the PCR re­ac­tion is des­troyed by poly­merase, which is ac­com­pan­ied by the re­lease of a dye, and the fluor­es­cence of the mix­ture in­creases. This fluor­es­cence is meas­ured by a Real-Time PCR sys­tem in par­al­lel on sev­eral chan­nels.

Amp­li­fic­a­tion res­ults are re­cor­ded us­ing the fol­low­ing fluor­es­cence de­tec­tion chan­nels:

«ALSENSE FLU» Reverse-Transcription PCR Kit (SF-090–96, for 96 samples):

• Reaction buffer – 1,4 ml × 1 tube;
• Enzyme mixture – 100 µl × 1 tube;
• Positive control – 100 µl × 1 tube;
• Negative control – 500 µl × 1 tube.