Faststart essential dna green master

The total RNA from each sample was isolated with Trizol reagent (Thermo Scientific, Rockford, IL, USA) according to the manufacturer’s protocol. cDNA synthesis was synthesized by reverse transcription using cDNA Synthesis Kit (Roche, Mannheim, Germany). Q-PCR was performed using FastStart Essential DNA Green Master (vazyme, China) on a Roche LightCycler 96 Real-Time PCR System and the following primers:

RAD51	Forward primer	5′-CAACCCATTTCACGGTTAGAGC-3’
	Reverse primer	5′-TTCTTTGGCGCATAGGCAACA-3’
MCM5	Forward primer	5′-ATGTCGGGATTCGACGATCCT-3’
	Reverse primer	5′-CCAGGTTGTAATGCCGCTTG-3’
CDC7	Forward primer	5′- GAGGCGTCTTTGGGGATTCAG-3’
	Reverse primer	5′-GGTCCTACTTGTAACTGTGCTG-3’
HK2	Forward primer	5′-GAGCCACCACTCACCCTACT-3’
	Reverse primer	5′-CCAGGCATTCGGCAATGTG-3’
LDHA	Forward primer	5′-ATGGCAACTCTAAAGGATCAGC-3’
	Reverse primer	5′-CCAACCCCAACAACTGTAATCT-3’
β-actin	Forward primer	5′-CATGTACGTTGCTATCCAGGC-3’
	Reverse primer	5′-CTCCTTAATGTCACGCACGAT-3’

For quantification of circSNRK, miR-103-3p, or SNRK mRNA, PrimeScript™RT reagent Kit with gDNA Eraser (Vazyme biotech, Nanjing, China) was used to synthesize the first-strand cDNA. Stem-loop qRT-PCR was performed using a FastStart Essential DNA Green Master (Vazyme biotech, Nanjing, China). GAPDH was used to normalize the expression of circSNRK and SNRK mRNA. For quantification of miR-103-3p, Bulge-loop^TM miRNA qRT-PCR Primer Sets (one RT primer and a pair of qPCR primers for each set) specific for miR-103-3p is designed by RiboBio (Guangzhou, China), cDNA was synthesized with a miRNA 1st Strand cDNA Synthesis Kit (by stem-loop) (Vazyme biotech, China). AceQ qPCR SYBR Green Master Mix (Vazyme biotech, Nanjing, China) was then used for real-time PCR. U6 was used to normalize the cellular miR-103-3p expression. The formula was: relative gene expression = 2^−ΔΔCt. The primers are listed in Supplementary Table 1.