We firstly homogenized the bone specimens with a high-speed agitation, Polytron blender (Kinematica, Luzern, Switzerland). To extract total RNA, RNAiso Plus reagent (Takara) was used for BMSCs lysates, MC3T3-E1 cell lysates, and bone specimens. Next, RNA RT reactions were conducted with a PrimeScript RT reagent Kit and a genomic DNA Eraser (Perfect Real Time) (Takara) following the manufacturer's instruction. In the end, quantitative real-time PCR was accomplished by using an SYBR Premix Ex Taq II kit (Takara). Primer sequences used for qPCR were provided in Supplementary Table 2. Small nuclear RNA U6 was selected as an internal reference of miR-146a-5p and the housekeeping gene Actin was selected as an internal reference of mRNA.
Genomic dna eraser perfect real time
Manufactured by Takara Bio
The Genomic DNA Eraser (Perfect Real Time) is a laboratory equipment designed for the efficient removal of genomic DNA from RNA samples. It is a crucial tool for researchers and scientists who require high-quality, DNA-free RNA for various applications, such as real-time PCR analysis and RNA-sequencing.
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Lab products found in correlation
Sybr premix ex taq 2 kit, by Takara Bio (1 mentions)
Rnaiso plus reagent, by Takara Bio (1 mentions)
Primescript rt reagent kit, by Takara Bio (1 mentions)
Polytron blender, by Kinematica (1 mentions)
Sybr premix ex taq tli rnaseh plus, by Takara Bio (1 mentions)
Primescript reverse transcriptase reagent kit, by Takara Bio (1 mentions)
2 protocols using genomic dna eraser perfect real time
1
Quantitative RNA Analysis Protocol
2
Quantifying RNA Expression in BE(2)-C Cells
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RNA was extracted from BE(2)-C cells using the RNeasy kit (Omega, USA). cDNA was produced using the PrimeScript Reverse Transcriptase reagent kit with genomic DNA Eraser (Perfect Real-Time) (Takara). qRT-PCR was performed using an Applied Biosystems 7900 HT qPCR machine and SYBR Premix Ex Taq (Tli RNaseH Plus) (Takara). The relative abundance of cDNA was calculated using the relative standard curve method.
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