Reference gene selection for miRNA and mRNA normalization in potato in response to potato virus Y.
Mol Cell Probes
; 55: 101691, 2021 02.
Article
in En
| MEDLINE
| ID: mdl-33358935
ABSTRACT
This was the first report on evaluating candidate reference genes for quantifying the expression profiles of both coding (e.g., mRNA) and non-coding (e.g., miRNA) genes in potato response to potato virus Y (PVY) inoculation. The reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) method was employed to quantify the expression profiles of eight selected candidate reference genes; their expression stability was analyzed by four statistical algorithms, i.e., geNorm, BestKeeper, NormFinder and RefFinder. The most stable reference genes were sEF1a, sTUBb and seIF5 with a high stability. The least stable ones were sPP2A, sSUI1 and sGAPDH. The same reference gene allows for normalization of both miRNA and mRNA levels from a single RNA sample using cDNAs synthesized in a single RT reaction, in which a stem-loop primer was used for miRNAs and the oligo (dT) for mRNAs.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Solanum tuberosum
/
RNA, Messenger
/
Genes, Plant
/
Potyvirus
/
MicroRNAs
/
Real-Time Polymerase Chain Reaction
Language:
En
Journal:
Mol Cell Probes
Journal subject:
BIOLOGIA MOLECULAR
/
BIOTECNOLOGIA
Year:
2021
Type:
Article