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Total RNA was extracted using RNeasy Mini Kit (Qiagen) or Tri Reagent (Sigma Aldrich) according to the manufacturer's instructions. Contaminating DNA was removed by DNA-free Kit DNA Removal Kit (Ambion). RNA was reverse transcribed using High Capacity cDNA Reverse Transcription Kit (Applied Biosystems). For Real time PCR, gene-specific primers for iNOS (20 (link)), Arg I (21 (link)), TNF-α (22 (link)), IL-10 (23 (link)), TGF-ß (24 (link)), MMP-2 (25 (link)), MMP-9 (26 (link)), Collagen-1, Collagen-3 (27 (link)), IL-6, 18S RNA (19 (link)), Fizz1/RELMα, Ym-1 (28 (link)), SPHK1, LIGHT (29 (link)), MertK (30 (link)), were used. All samples were run in triplicates. Relative gene expression levels were determined using PowerUP SYBR Green PCR Master Mix (Applied Biosystems) according to the manufacturer's recommended protocol with the following thermal cycling conditions: 2 min 50°C, 2 min 95°C, 40 cycles of 1 s 95°C and 30 s 60°C, and 4°C hold (QuantStudio 3 Real-Time PCR System, Applied Biosystems). Expression of target genes was normalized to the endogenous control 18S RNA gene. Fold expression was calculated using the 2^−ΔΔCt methods. In some experiments, the 2^ΔCt method was used to determine relative gene expression.

Total RNA was extracted using RNeasy Mini Kit (Qiagen) according to the manufacturer’s instructions. Contaminating DNA was removed by DNA-free Kit DNA Removal Kit (Ambion). RNA was reverse transcribed using High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems). For real-time PCR, gene-specific primers for iNOS [16 (link)], Arg I [17 (link)], COX-2 [18 (link)], TNF-α [19 (link)], Fizz1/RELMα, Ym-1 [20 (link)], SPHK1, LIGHT [21 (link)], MertK [22 (link)], IFN-ɣ, IL-2, IL-4 [23 (link)], IL-10 [24 (link)], and GATA-3 [25 (link)] were used. To detect IL-6 and 18S RNA expression, the following primers were used: IL-6 forward 5′-CCACTTCACAAGTCGGAGGCTTA-3′; IL-6 reverse 5′-GCAAGTGCATCATCGTTGTTCATAC-3′; 18S RNA forward 5′-CGGCTACCACATCCAAGGAA-3′, 18S RNA reverse 5′-GCTGGAATTACCGCGGCT-3′.
All samples were run in triplicates. Relative gene expression levels were determined using Power SYBR Green PCR Master Mix (Applied Biosystems) according to the manufacturer’s recommended protocol with following thermal cycling conditions: 10 min 95 °C, 40 cycles of 15 s 95 °C and 60 s 60 °C, and 4 °C hold (StepOnePlus Real-Time PCR System, Applied Biosystems). Expression of target genes was normalized to the endogenous control 18S RNA gene. Fold expression was calculated using the 2^−ΔΔCT method [26 (link)].

Total RNA was extracted using RNeasy Mini Kit (Qiagen) according to the manufacturer’s instructions. Contaminating DNA was removed by DNA-free Kit DNA Removal Kit (Ambion). RNA was reverse transcribed using High Capacity cDNA Reverse Transkription Kit (Applied Biosystems). Primer sequences used for Real time PCR are listed in Table 1. All samples were run in triplicates. Relative gene expression levels were determined using Power SYBR Green PCR Master Mix (Applied Biosystems) according to the manufacturer’s recommended protocol with following thermal cycling conditions: 2 min 50°C, 2 min 95°C, 40 cycles of 1 sec 95°C and 30 sec 60°C, and 4°C hold (QuantStudio 3 Real-Time PCR System, Applied Biosystems). Expression of target genes was normalized to the endogenous control 18S RNA gene. Fold expression was calculated using the 2^-ΔΔCT method. In some experiments, the 2^ΔCT method was used to determine relative gene expression.