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. 2022 Aug 2;23(15):8590. doi: 10.3390/ijms23158590

Table 3.

DNA-based vs. RNA-based approaches, choosing the right starting material for TCR profiling.

Advantages
gDNA mRNA
  • easier to obtain;

  • very stable [49];

  • no requirement for reverse transcription (RT);

  • better reflect the number of analyzed cells;

  • accurate measurement of clonality without bias caused by variable expression levels in different cells.

  • higher number of copies in a single cell;

  • large information at the gene transcription level;

  • reduced interference of non-coding signals after the splicing process [50];

  • overall length sequence in the CDR region is easily available;

  • non-productive receptor transcripts are underrepresented [51].

  • close proximity of V and C regions after the splicing process facilitates PCR amplification [13].

Disadvantages
gDNA mRNA
  • higher concentration input;

  • potential annealing of primers for multiple binding sites;

  • presence of introns and “unused” segments in the sequence of interest that have to be amplified, causing challenges during PCR process [13];

  • Detection of all the TCR sequences whether they contribute to a productive or a nonproductive segment arrangement [13].

  • introduction of errors during retrotranscription [52];

  • easily degraded;

  • high requirements for extraction, transportation and storage.