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. Author manuscript; available in PMC: 2020 Apr 15.
Published in final edited form as: Methods. 2019 Jan 24;159-160:129–137. doi: 10.1016/j.ymeth.2019.01.010

Fig. 4.

Fig. 4.

Testing the functionality of UAS-Rpb1 derivatives using the Rpb1G0040 allele. (A) Schematic of the Rpb1G0040 rescue assay. “C” type males have normal body color due to y+ while “B” type males have yellow body color due to y−. (B) Schematic for the PCR evaluation of the endogenous Rpb1 allele located on the X chromosome in progeny from the Rpb1G0040 rescue assay. Transcription start site (TSS) of Rpb1 is shown. Blue arrows represent primers. PCR amplification of the wild-type endogenous allele with primers F and R1 produces a 205 bp DNA. PCR amplification with primers F and R2 of the Rpb1G0040 allele produces a 577 bp DNA. Sequences of primers: F: 5’-AGAAGGCTGGGTAAACAATCAC-3’, R1: 5’-CTGCTACAACGACCGCAATA-3’ and R2: 5’-AATGAACAGGACCTAACGCACA-3’. (C) PCR results for the endogenous Rpb1 allele following the scheme shown in (B). Genomic DNA was isolated from one male fly [34] and amplified with a mixture of primers F, R1 and R2. Lane 1 shows the product from one male containing the endogenous wild-type allele but not the Rpb1G0040 allele. Consequently, only the 205 bp DNA is produced. Lane 2 shows the product from a “C” male. The presence of a 577 bp product confirms the presence of Rpb1G0040 while the absence of the 205 bp product indicates the absence of the endogenous wild-type allele.