In brief, PCR reaction was performed in 20 μl containing 100 ng of template DNA, 4 μL of × 5 PCR buffer, 0.7 μL (10 pmol/μL) of each primer,[13 (link)] 1.6 μL of 10 mM deoxynucleoside triphosphates, and 0.4 μL of GXL Taq polymerase (Takara, Clontech). The PCR conditions were: 95°C for 15 min followed by 40 cycles of 95°C for 30 s, 56°C for 1 min, 72°C for 1 min, and final extension at 72°C for 10 min. PCR products were analyzed in 1.5% agarose gel and subjected to purification using EXO-SAP IT (USB, Affymetrix).
Gxl taq polymerase
Manufactured by Takara Bio
GXL Taq polymerase is a high-fidelity DNA polymerase designed for accurate DNA amplification. It exhibits enhanced processivity and thermal stability, enabling efficient and reliable PCR performance.
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3 protocols using gxl taq polymerase
1
PCR Amplification and Purification Protocol
2
Mouse Cardiac cDNA Sequencing
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The full-length cDNA sequence of Cfast was obtained from mouse hearts RNA by applying RACE according to SMARTer RACE 5′/3′ Kit User Manual (TaKaRa). Nested 5′ and 3′ RACE products were obtained using GXL Taq polymerase with GC Buffer (Takara). The primers used for RACE, nested PCR, and PCR are presented in Table S4 . Gel products were extracted with a Gel Extraction kit (Vazyme Biotech), cloned into pMD18-T vectors and analyzed by Sanger sequencing.
3
Full-Length cDNA Sequencing of LncHrt
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The full-length cDNA sequence of LncHrt was obtained from mouse heart RNA according to the SMARTer RACE 5ʹ/3ʹ Kit User Manual (TaKaRa). Nested 5ʹ and 3ʹ RACE products were obtained using GXL Taq polymerase with GC Buffer (Takara). The primers used for RACE PCR and nested PCR are presented in Supplementary Table 12. PCR products were extracted with a Gel Extraction kit (Vazyme Biotech), cloned into the pMD18-T vector, and analyzed by Sanger sequencing.
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