Fig. 1
From: Degenerate codon mixing for PCR-based manipulation of highly repetitive sequences
Generating the putative Q52, Q31 and Q10-GFP constructs using the Tol2-Q80-GFP-v2A-GFP multicistronic reporter construct. a Vector map of the Tol2-Q80-GFP-v2A-GFP construct. b Summary of CAA and CAG degenerate codon usage in this construct. c Chromatograph of the construct depicting the randomly interspaced glutamine coding CAG and CAA triplets. d Schematic illustration of PCR-based exclusion amplification of the Q80-GFP containing construct to generate polyQ constructs with lower numbers of glutamine repeats. e Q80 sequence showing Q52, Q31 and Q10 primer binding sites. f Sequences of primers intended to generate Q52, Q31 and Q10 constructs. g Analytical agarose gel electrophoresis for each of the putative Q80, Q52, Q31 and Q10 vectors using primers flanking the polyQ region. PCR product sizes of ~ 270, ~ 180, ~ 120 and ~ 60 bp for the intended putative Q80, Q52, Q31 and Q10-GFP constructs, respectively, were seen