Cloning Repebody pET303-EGFP-HA Link to PDF pET303-mCherry-HA Link to PDF pET303-unmod-Repebody-6xHis Link to PDF pET303-ssDNA-optimized-Repebody-6xHis Link to PDF pCDFDuet-unmod-Repebody-6xHis Link to PDF pCDFDuet-ssDNA-Repebody-6xHis Link to PDF pCDFDuet-unmod-Repebody-6xHis-EGFP-HA Link to PDF pCDFDuet-ssDNA-Repebody-6xHis-EGFP-HA Link to PDF pCDFDuet-unmod-Repebody-6xHis-mCherry-HA Link to PDF pCDFDuet-ssDNA-Repebody-6xHis-mCherry-HA Link to PDF Retron pMS366-ret-rb-rs-gb-single Link to PDF pMS366-ret-rb-rs-gb-single-ret-rb-rs-gb-double Link to PDF pMS366-ret-rb-rs-gb-single-ret-rb-rs-gb-double-fixed Link to PDF pMS366-Ret & pMS366-RibojRet Link to PDF pJet-Ret & pJet-RibojRet Link to PDF pET303-EGFP restriction site removal SDM Link to PDF pET303-mCherry restriction site removal SDM Link to PDF pET303-EGFP-G92D SDM Link to PDF pET303-EGFP-G68P SDM Link to PDF pET303-EGFP-G21D SDM Link to PDF pCDFDuetKO Link to PDF pCDFDuetIso Link to PDF pCDFDuetIso-Repebody Link to PDF pCDFDuetIso-mCherry Link to PDF pCDFDuet1-mCherry Link to PDF pCDFDuetIso-EGFP-Repebody Link to PDF pCDFDuetIso-EGFP-mCherry Link to PDF pCDFDuet1-EGFP-mCherry Link to PDF Selection pKNT25-unmodified-mCherry-binding Repebody-6xHis Link to PDF pKNT25-unmodified-mCherry-binding Repebody-6xHis-GGS Link to PDF pKNT25-ssDNA-optimized-mCherry-binding Repebody-6xHis Link to PDF pKNT25-ssDNA-optimized-mCherry-binding Repebody-6xHis-GGS Link to PDF pKT25-unmodified-mCherry-binding Repebody-6xHis Link to PDF pKT25-unmodified-mCherry-binding Repebody-6xHis-GGS Link to PDF pKT25-ssDNA-optimized-mCherry-binding Repebody-6xHis Link to PDF pKT25-ssDNA-optimized-mCherry-binding Repebody-6xHis-GGS Link to PDF pUT18-eGFP-HA Link to PDF pUT18-eGFP-HA-GGS Link to PDF pUT18-mCherry-HA Link to PDF pUT18-mCherry-HA-GGS Link to PDF pUT18C-eGFP-HA Link to PDF pUT18C-eGFP-HA-GGS Link to PDF pUT18C-mCherry-HA Link to PDF pUT18C-mCherry-HA-GGS Link to PDF TPD pET303-p62-GGSGG-10xHis Link to PDF pET303-GBP-linker-10xHis Link to PDF pET303-mGBP-linker-10xHis Link to PDF pET303-A1E-linker-HA Link to PDF pcDNA3-1-EGFP-mCH-FIS Link to PDF pcDNA3-1-mCH-EGFP-FIS Link to PDF pcDNA3-1-GBP-mTagBFP2-HA-A1E Link to PDF pcDNA3-1-mGBP-mTagBFP2-HA-A1E Link to PDF pcDNA-rb-BFP-HA Link to PDF pcDNA-rb-BFP-HA-A1E Link to PDF Repebody characterization Test - unmod-Repebody: expression and binding Link to PDF 800 ml cultures of unmod- Repebody & fluorescent proteins for gelfiltration & concentration measurement Link to PDF ssDNA-expression 1L culture Link to PDF Test - ssDNA-Repebody: expression in pET303 Link to PDF Test - ssDNA-Repebody: expression in pCDF-DuetIso Link to PDF Quantitative ssDNA-Repebody Pulldown with normalized mCherry-HA to EGFP-HA concentrations Link to PDF Far Western Blot of mCherry-HA and EGFP-HA with ssDNA-Repebody as staining solution Link to PDF Test - unmodified-Repebody & ssDNA-Repebody: expression in pET303 Link to PDF Repebody Coexpression pulldown Link to PDF Western Blot: mCherry-HA & EGFP-HA Link to PDF Western Blot: mCherry-HA & EGFP-HA (2) Link to PDF IP- Binding assay: Unmodified Repebody to mCherry & EGFP Link to PDF IP- Binding assay: Unmodified Repebody to mCherry & EGFP (2) Link to PDF IP- Binding assay: ssDNA Repebody & Unmodified Repebody to mCherry & EGFP Link to PDF Retron qPcr:pms366. determine ideal temperature Link to PDF qPcr: pms_366. determine standard curve (1) Link to PDF qPcr: pms_366. determine standard curve (2) Link to PDF qPcr: pms_366. determine standard curve (3) Link to PDF qPcr: pms_366. quantify editing rates (1) Link to PDF qPcr: pms_366. quantify editing rates (2) Link to PDF qPcr: pms_366:Retron. determine standard curve Link to PDF qPcr: pms_366:Riboj-Retron. determine standrd curve Link to PDF pcr test for pcdfduetiso:rb primers Link to PDF pcr test for pcdfduetiso:rb primers qualitatively test retron integration I - minprepped sample Link to PDF EGFP-mCherry counterfluorescence expression (1) Link to PDF EGFP-mCherry counterfluorescence expression (2) Link to PDF EGFP Loss-of-Function mutant screening Link to PDF pCDFDuetIso-Repebody expression, 18˚C, 25˚C (1) Link to PDF pCDFDuetIso-Repebody expression, 18˚C, 25˚C (2) Link to PDF pCDFDuetIso-Repebody expression, 30˚C Link to PDF Selection BACTH - qualitative assay Link to PDF BACTH - dot-blot of samples 6-8 Link to PDF BACTH - serial dilution of samples 6-14 Link to PDF TPD Pulldown: EGFP-GBP Link to PDF Pulldown: pET303-(m)GBP-10xHis & EGFP-HA / mCherry-HA Link to PDF Pulldown: pET303-A1E-linker-HA binding to pET303-p62-linker-10xHis & pET303-GBP-linker-10xHis binding to pET303-EGFP-HA Link to PDF Pulldown: pET303-A1E-linker-HA & pET303-p62-linker-10xHis Link to PDF p62-mCherry, EGFP in U2OS Link to PDF Confocal microscopy: Co-transfection of EGFP, p62-mCherry, mTagBFP2 Link to PDF PEI transfection optimisation in U2OS cells Link to PDF FACS: EGFP mCherry mTagBFP2 in U2OS Link to PDF FACS: EGFP mCherry mTagBFP2 transfection in U2OS TRex Link to PDF PEI transfection optimisation in HeLA cells Link to PDF FACS: Transfection optimisation in HeLa cells Link to PDF FACS #2: Transfection optimisation in HeLa cells Link to PDF Confocal Microscopy: mitoEGFP; EGFP-mCherry Transfection under EBSS Starvation in HeLA cells Link to PDF Confocal Microscopy 1#: Single transfection TARGETS Link to PDF Western blot of the expression of target (EGFP-mCherry, EGFP-mCh-FIS, mCherry-EGFPFIS) and RB-construct in HELA cell Link to PDF FACS: Targets EGFP-mCherry, EGFP-mCherry-FIS, mCherry-EGFP-FIS in HeLa cells Link to PDF Confocal Microscopy: Single transfection DEGRADERS Link to PDF FACS/WB: Repebody degradation + EBSS treatment Link to PDF Western Blot: p62mCherry, RB deg Degradation Link to PDF Confocal Microscopy 2# : Single transfection TARGETS Link to PDF
