The procedure of transcription based on AFLP grading method

The procedure of transcription based on AFLP grading method

AFLP procedure is carried out in a few stages. First of all, the common RNA is separated. Afterwards poli-A is separated and synthesis of double-chain cDNA is performed, at the next stage it is affected by two different restriction fragments. These fragments cut cDNA into fragments of 50 – 800 bp size. Since the restriction link is included into oligo(dT)25V primer, 5’-cDNA fragments are eliminated even before the inclusion of the second restriction enzyme into the reaction. Then anchors are attached to the cut fragments at the point of the start of their transcription and PCR is performed. This reaction employs primers, complying with the sequence of the anchors. In the further amplification reaction, the cut fragments are amplified using special cDNA 3’-end primers. At the last stage one-chained DNA fragments are distributed applying electrophoresis method and depending on the way they were marked, observed using autoradiograph, painting or with the help of flourescence.

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Figure 5. Scheme of differentiation regulated by differentiation