Takara rna pcr kit

Total RNA was extracted from the liver tissues with RNAiso Plus reagent (TaKaRa Bio, Shiga, Japan). The cDNAs were synthesized using the AMV reverse transcriptase and random 9-mers in the TaKaRa RNA PCR Kit (version 3.0; TaKaRa Bio). The primer sets used for PCR amplification are listed in Table S1 (Supplementary Materials). Quantitative reverse-transcription PCR was performed with SYBR Green (TaKaRa), using a TaKaRa TP-815 instrument. The relative quantities of amplified DNA were analyzed using the software bundled with the TP-815 instrument and normalized to the mouse 36B4 mRNA level.

Total RNA of PC-3, DU145, LNCaP, 22RV1, RWPE-1 or BPH1 cells after various treatments was extracted with TRIzol-reagent (Tel-Test; Austin, Texas, USA). First-strand cDNA was synthesized by reverse transcription of 2 μg total RNA using a Takara RNA PCR Kit (TaKaRa, Japan). The primers used were: glyceraldehyde 3-phosphase dehydrogenase (GAPDH): Hs02758991; HIF-1α: Hs00153153; HK2: Hs00606086; PDK1: Hs00176853; PKM: Hs00761782; and LDH-A: Hs01378790 (Invitrogen). The expression levels of target genes were normalized to that of the housekeeping gene GAPDH.

The total RNA from hippocampus and cortex tissues prepared in advance was extracted using TRIZOL reagent (Invitrogen, USA). Then 2 μg of RNA of each sample was transcribed to cDNA using the TaKaRa RNA PCR™ kit (Takara, Japan). The mRNA expression of Bdnf, Sirt1, App, Bace1 and Gadph was inspected using cDNA as a template for amplification utilizing the following primers: Bdnf IV (forward primer 5'-CAGAGCAGCTGCCTTGATGTT-3' and reverse primer 5'-GCCTTGTCCGTGGACGTTTA-3'); Sirt1 (forward primer 5'-GAGGTCTCGA TATGTGCTGGA-3' and reverse primer 5'-TTCCTGCAACCTGCTCCAAG-3'); App (forward primer 5'-GACTGACCACTCGACCAGGTTCTG-3' and reverse primer 5'-CTTGTAAGTTGGATTCTCATATCCG-3'); Bace1 (forward primer 5'-CCTACACCCAGGGCAAGT-3' and reverse primer 5'-GGGCAGTAGTAAC TTTGCAGT-3'); and Gapdh (forward primer 5'-CTTCCAGGAGCGAGACC-3' and reverse primer 5'-CGGAGATGATGACCCTTTT-3'). The expression levels of each sample were normalized against Gadph (internal control) and calculated using the comparative CT method (2^−ΔΔCT).

Total RNA was extracted from cultured B-ALL cells using TRIzol (Invitrogen, Waltham, MA, USA), following the manufacturer’s instructions. The RNA concentration was examined by a NanoDrop™ 2000UV–Vis Spectrophotometer (ThermoFisher, Waltham, MA, USA). Complementary DNA (cDNA) were produced by using the TaKaRa RNA PCR Kit (TaKaRa Bio, Kusatsu, Shiga, Japan). The cDNA was amplified by SYBR Green real-time qPCR Kit (TaKaRa Bio) according to the manufacturer’s instructions. The expression levels of mRNA were measured by 2^−ΔCt method. β-actin expression levels were used as internal control. The primers used in real-time (qRT-PCR) are shown in Additional file 1: Table S1. Each assay was performed in triplicate.