Fast sybr green quantitative pcr kit

Total RNA was prepared from cells using the RNA extraction kit, RNeasy Mini Kit (Qiagen, Hilden, Germany) as per the manufacturer’s instructions. Of total RNA, 0.5 or 1 μg was converted into single-strand cDNA using Superscript II (Invitrogen, Carlsbad, CA, USA) as per the manufacturer’s instructions. RT-qPCR was performed with Fast SYBR^®Green quantitative PCR kit (Applied Biosystems, Foster City, CA, USA) for RPL19 (Forward primer sequence: 5′-CGGAAGGGCAGGCACAT-3′ and Reverse primer sequence 5′-GGCGCAAAATCCTCATTCTC-3′), IL-8 (Forward primer sequence: 5′-AGGGTTGCCAGATGCAATAC-3′ and Reverse primer sequence 3′-CCTTGGCCTCAATTTTGCTA-5′) and PTEN (Forward primer sequence: 5′-AATAAAGACAAAGCCAACCGATACTT-3’ and Reverse primer sequence 5’-CGGCTCCTCTACTGTTTTTGTGA-3′). Expression of PTEN and IL-8 mRNA was then normalized with RPL19 and PTEN was compared with mRNA positive control of T98G.

Total RNA was prepared from cells using the RNA extraction kit, RNeasy Mini Kit (Qiagen, Hilden, Germany) as per the manufacturer’s instructions. Of total RNA, 1 μg was converted into single-strand cDNA using Superscript II (Invitrogen, Carlsbad, CA, USA) as per the manufacturer’s instructions. Quantitative real-time PCR (RT-qPCR) was performed with Fast SYBR^®Green quantitative PCR kit (Applied Biosystems, Foster City, CA, USA) for RPL19 (Forward primer sequence: 5′-CGGAAGGGCAGGCACAT-3′ and Reverse primer sequence 5′-GGCGCAAAATCCTCATTCTC-3′) and PIK3CD (Forward primer sequence: 5′-CCCACATGAAGAGGAACTGAGAT-3′ and Reverse primer sequence 3′- GGTTGGCAGGCTCAGTGACT-5′). Expression of PIK3CD was then normalized with RPL19.