Abi550 real time pcr machine

RNA isolation and RT-qPCR were performed as described [40 (link)]. The RT-qPCR tests were performed using an ABI550 Real-Time PCR machine (Applied Biosystems, Thermo Fisher Scientific, Austin, TX, USA) and the 2x SYBR Green qPCR Mix (Aidlab, Beijing, China). The primers used for verification of the effects of silencing of GmSAUL1a/1b, were:
GmSAUL1a/1b-V-F: ATGATGGCTGCGAGCT;
GmSAUL1a/1b-V-R: CTCATGTGAAAGGAATTTTACTAC.
The additional primers used in this study were:
GmSAUL1a/1b-F: ATGATGGCTGCGAGCT;
GmSAUL1a/1b-R: CTCATGTGAAAGGAATTTTACTAC;
GmELF1b-F: ACCGAAGAGGGCATCAAATCCC;
GmELF1b-R: CTCAACTGTCAAGCGTTCCTC;
GmELF1b-F (RT-qPCR): GTTGAAAAGCCAGGGGACA;
GmELF1b-R (RT-qPCR): TCTTACCCCTTGAGCGTGG;
GmPR1-F (RT-qPCR): ATGGGGTTGTGCAAGGTT;
GmPR1-R (RT-qPCR): CTAGTAGGGTCTTTGGCCAA.

Total RNA was extracted with Trizol (Invitrogen, Waltham, MA, USA). RT-qPCR was performed as described [22 (link)] using the 2× SYBR Green qPCR Mix (Aidlab, Beijing, China) and an ABI550 Real-Time PCR machine (Applied Biosystems, Thermo Fisher Scientific, Austin, TX, USA). The following primers were used:
qRT-NtUBQ5-F: TCTTCATCTCGTGCTCCG;
qRT-NtUBQ5-R: TCCAGAATCATCCACCTTGTA;
qRT-NtPR1-F: GCTGAGGGAAGTGGCGATTTC;
qRT-NtPR1-R: CCTAGCACATCCAACACGAACC;
qRT-NtPR3-F: GACCCATCCAATTGACAAACCAA;
qRT-NtPR3-R: CTGTGGTGTCATCCAGAACC;
qRT-NtPR5-F: GGCGATTGTGGCTCAAACC;
qRT-NtPR5-R: GAAATCTTGCTTCGTACCTGAGA.