Primescript rt reagent with gdna erase

Total RNA of all tissues was purified with TRIzol (15596026; Invitrogen) and converted to cDNA with a PrimeScript™ II 1st Strand cDNA Synthesis Kit (No.6210A; Takara, Dalian, Liaoning Province, China) according to the manufacturer's protocol. qPCR analysis was conducted with PrimeScript™ RT reagent with gDNA Erase (No. RR047Q; Takara), as previously described 19. Constructs that expressed Odf1/2/3/4 and Cenexin‐GFP fusion proteins were cloned from adult testicular cDNA. The primers used in these assays are listed in Table S1.

Total RNA was extracted from tissues with TRIzol (15596026, Invitrogen, Carlsbad, CA, U.S.A.) and converted to cDNA using the PrimeScript™ II First Strand cDNA Synthesis Kit (No. 6210A, Takara, Japan). The Cfap58 cDNA fragment was amplified from adult testis cDNA library and cloned into a pcDNA3.0 empty vector digested by EcoR V and Xho I. For the qPCR assay, the samples were conducted with PrimeScript™ RT reagent with gDNA Erase (No. RR047Q, Takara, Japan) as previously described [28 (link)]. The RNAi-resistant Cfap58 against Cfap58 sh3 was performed by PCR-based mutagenesis. The mutant allele contains 5′-gAatgtcCatgaaTaaTatTt-3′ (from nt2522 to nt2542 in CDS) (mutant alleles are indicated in uppercase). The primers used in this section are listed in Supplementary Table S2.
The shRNA sequences targeted mouse Cfap58 were designed using an online design tool in Thermo Fisher Scientific and cloned into a pLKD vector under the control of the U6 promoter. The sequences were given as following Supplementary Table S2.