Quant one step qrt pcr

HSV-1 infected and control mice were euthanized. The brain tissues were harvested and homogenated in Trizol (Transgen China, ET101-01). RNA was extracted according to manufacturer protocol and subjected to Real-time qPCR analyses with Quant One Step qRT-PCR (Tiangen, China) on 7500 Real Time PCR System (Applied Biosystems, USA). Relative quantification of HSV-1 gB mRNA was determined by using GAPDH as internal control and 2^−ΔΔCt method. The primer sequences were as follows: HSV-1 gB forward: 5′-AACGCGACGCACATCAAG-3′; HSV-1 gB reverse: 5′-CTGGTACGCGATCAGAAAGC-3′; GAPDH forward: 5′-GCATTGTGGAAGGGCT CA-3′; GAPDH reverse: 5′- ACCAGTGGATGCAGGGAT-3′; Uchl1 forward: 5′-AGGGACAGGAAGTTAGCCCTA-3′; Uchl1 reverse: 5′-AGCTTCTCCGTTTCAGA CAGA-3′; Icam5 forward: 5′-TCCGAACTTTCCAGCGACC-3′; Icam5 reverse: 5′-CTACGAAACTGCGGCGAATC-3′; Hcrt forward: 5′-GTCGCCAGAAGACGTGTT C-3′; Hcrt reverse: 5′-GTGGTAGTTACGGTCGGACA-3′; Eif2ak2 forward: 5′-ATGCACGGAGTAGCCATTACG-3′; Eif2ak2 reverse: 5′-TGACAATCCACCTTGTTTTCGT-3′; Oas1a forward: 5′-GCCTGATCCCAGAATCTATGC-3′; Oas1a reverse: 5′-GA GCAACTCTAGGGCGTACTG-3′; Oas2 forward: 5′-TTGAAGAGGAATACATGCG GAAG-3′; Oas2 reverse: 5′-GGGTCTGCATTACTGGCACTT-3′; Oas3 forward: 5′-T CTGGGGTCGCTAAACATCAC-3′; Oas3 reverse: 5′-GGCAATCCTTATCACTCTT GGTC-3′.