iscripttm gdna clear cdna synthesis kit bio rad  (Bio-Rad)

Journal: Frontiers in Neurology

Article Title: A novel truncating variant of SPAST associated with hereditary spastic paraplegia indicates a haploinsufficiency pathogenic mechanism

doi: 10.3389/fneur.2022.1005544

Figure Lengend Snippet: Transcription analysis of spastin in lymphoblast cell lines. (A) Agarose gel electrophoresis of the long-range RT-PCR products revealed only the cDNA fragment of 1,494 bp corresponding to the predicted canonical transcripts in both the patient and the control. (B) Sanger sequencing of long-range RT-PCR products revealed only the canonical transcript in the proband, which was reconfirmed on sequencing with a reverse primer. The transcript with the frameshift variant was not detected. (C) Agarose gel electrophoresis of the shorter RT-PCR products revealed only the 603 bp band expected in both the patient and control. (D) Sanger sequencing of the shorter RT-PCR products in both directions identified only canonical transcripts in the patient. cDNA, complementary DNA; gDNA, genomic DNA.

Article Snippet: Reverse transcription was performed using a Bio-Rad's iScriptTM gDNA Clear cDNA Synthesis Kit (Bio-Rad Laboratories, Hercules, CA).

Techniques: Agarose Gel Electrophoresis, Reverse Transcription Polymerase Chain Reaction, Control, Sequencing, Variant Assay