Sybr green real time pcr master mix plus kits

Total RNAs used in the present study were extracted from the tissues using the TRIzol reagent (Invitrogen, Carlsbad, CA). The cDNA synthesis was performed with 1 μg of total RNA according to the manufacturer’s protocol (Takara, Tokyo, Japan). GAPDH (forward, 5′-TCGGAGTCAACGGATTTGG-3′ and reverse, 5′-CATGGGTGGAATCATATTGGA-3′) was used as internal control in SYBR Green Real-time PCR Master Mix-Plus kits (Toyobo, Osaka, Japan). The relative RNA levels of GAS5 were determined with the Quant Studio 6 Flex system (Applied Biosystems, Life Technologies, U.S.A.). The relative quantitative value was shown by the 2^−ΔΔCt. The experiment was carried out in triplicates.

Total RNA samples from CRC cells or frozen CRC tissues were extracted using TRIzol reagent (Invitrogen). After that, 1 µg RNA/sample was reversed‐transcribed into first‐strand cDNA with PrimeScript™ RT Reagent Kit (Takara). SYBR Green Real‐time PCR Master Mix‐Plus kits (Toyobo) was used to perform qPCR in triplicate to test MAGI2‐AS3 and GR mRNA level. 2^‐ΔΔCt method was used for relative expression calculation. The experiment was carried out in triplicates. Primers used were listed here:
MAGI2‐AS3: 5′‐ATACAAGCCCAAGTTCTG‐3′(sense);
MAGI2‐AS3: 5′‐TTCCTGGTGTTTCCTCTT‐3′ (antisense);
GR: 5′‐TGCGTCTTCACCCTCACT‐3′ (sense);
GR: 5′‐CCAGGTCATTTCCCATCA‐3′ (antisense);
β‐actin: 5′‐ATCCGCAAAGACCTGT‐3′ (sense);
β‐actin: 5′‐GGGTGTAACGCAACTAAG‐3′ (antisense);

Total RNAs were extracted from frozen tissue samples of patients with cervical cancer using Trizol reagent (Invitrogen, US) and treated by DNase / RNase-free Deionized water (Tiangen, Beijing, China). cycA was used as internal controls in SYBR^® Green Realtime PCR Master Mix-Plus kits (Toyobo, Osaka, Japan). The expression level of XRCC1 were amplified with PCR primers (Takara, Shanghai, China) and quantitative using the Quant Studio 6 Flex system (Applied Biosystems, Life Technologies, USA).