Single Cell Transcriptome Amplification with MALBAC (original) (raw)
Fig 1
Single-cell MALBAC-RNA amplification diagram.
After reverse transcription, primers with 7 random nucleotides at the 3’ end are annealed to the cDNA template at 4°C, then extended by DNA polymerase with strand displacement activity as temperature is increased. Amplicons are then melted off the original template after DNA extension, and looped at 58°C to protect themselves from being further amplified thanks to their 5’ ends being complementary to their 3’ ends. This MALBAC-RNA step includes a total of 10 cycles of quasilinear amplification, followed by another 19 cycles of PCR.