Quantstudio 5 design and analysis software v1

The total RNA from cells was isolated with Ultraspec or TRIsure (BIO-38032, Bioline USA, Inc., Memphis, TN, USA) and this RNA (1 μg) was then reverse-transcribed using Improm-II reverse transcriptase (Promega, Madison, WI, USA). Polymerase chain reaction (PCR) amplifications were performed using the Power SYBR Green PCR Master Mix kit (Applied Biosystems, Foster City, CA, USA) in a QuantStudio5 (Applied Biosystems, Foster City, CA, USA) with the primer sets indicated below. The data were analyzed with QuantStudio5 Design and Analysis Software v1.4 (Applied Biosystems, Foster City, CA, USA). The following set of primers were used: Epo (forward 5’ -CAAAGTAACTTCTATGCTTGGAAAA-3′ reverse, 5′-CAGGCCTTGCCAAACTTCTATG-3′); Slc7a5 (forward 5′-TTCGCCACCTACTTGCTCAA-3′, reverse 5′-CCTTTACGCTGTAGCAGTTC-3′); 28S (forward 5′-GGTAGCCAAATGCCTCGTCAT-3′, reverse 5′-GGATAGTAGGTAGGGACAGTGGGAAT-3′). For amplification of murine genomic region lacking Slc7a5 exon1 in ErGFPcre Slc7a5^LoxP/LoxP mice the following primers were used forward 5′- GGCTCCTGGACTTATCTTGACCAATG-3′ reverse, 5′- GTGGTGCTTTGCTGAAGGCAGGG-3′).¹⁹ (link)