... you casually mention “Haery et al., showed that MYC expression was increased…” when discussing the review...

...target. In cases such as this, an epitope such as c-Myc, GFP or V5 can be used to tag the protein of interest...

...crRNA and/or try out an alternative tag (3xFLAG, Myc, V5, or HA). Tag-specific PCR genotyping can tell...

...expression ~ ~ ~ ~ ✓   ✓ SV40 and c-Myc nuclear localization sequences Improve translocation...

...while there are many types of epitope tags (eg. HA, MYC, Flag), they can be deleterious and cause aberrant...them to popularly used epitopes tags such as HA and MYC in vivo. The authors found that fusions to one particular...fragment with FKBP rapamycin binding domain of mTor (Cas9(N)-FRB-NES) resulting in a rapamycin-inducible Cas9...Addgene: Rapamycin-inducible Cas9 sets (Addgene plasmids 62883 &62884; 62885 & 62886) Rapamycin-inducible...separate reporter gene (EGFP, iRFP, IFP1.4, puromycin, neomycin or luciferase) to create a final lentiviral...Cas9 for genome editing. Without rapamycin treatment, the Cas9(N)-FRB-NES fragment is actively shuttled...to the nuclear export sequence. Treatment with rapamycin induces Cas9(N)-FRB-NES and Cas9(C)-FKBP-2xNLS...

...deposited HA tagged BioID2 for N-terminal fusions and Myc tagged BioID2 for C-terminal fusions. Kim, et al...pharmaceutical industry, depend on the yeast, Saccharomyces cerevisiae, as a key host for the production...designed to complement unused auxotrophies in Saccharomyces strains by compensating for histidine (HIS3)...

...Slow transformation   Spectinomycin/streptomycin Spectinomycin and streptomycin both inhibit protein synthesis.... Pros: Stable (spectinomycin). Cons: Cost (spectinomycin) or stability (streptomycin), doesn’t work for... for transformations. Con: Expensive.   Kanamycin Kanamycin, or kan, is an aminoglycoside which inhibits...Like carb, spectinomycin is the more stable and expensive antibiotic compared to streptomycin. Spectinomycin...Spectinomycin/streptomycin can be successfully used for plant and bacterial culture, but some types of bacteria...bacteria.   Zeocin  Zeocin is a member of the bleomycin family of antibiotics, which induce cell death...decorative" title="Check icon" %} Satellite colonies Kanamycin Excellent No Bacteria and eukaryotes (G418) {%...

...shorter plasmid curing time. pCRISPomyces for editing in Streptomyces  Streptomyces bacteria produce a wide...created two “pCRISPomyces” systems for use in Streptomyces (Cobb et al., 2015). pCRISPomyces-1 includes ...array, while pCRISPomyces-2 contains Cas9 and a gRNA cassette. The simpler system of pCRISPomyces-2 displays...for transcriptional silencing Mycobacterium tuberculosis and Mycobacterium smegmatis. In addition to their...carries a spacer targeting the gene of interest and kanamycin resistance E. coli carrying the phage recombineering...Gibson Assembly or restriction enzyme cloning. Streptomyces bacteria are more recombinogenic than E. coli...efficiencies or can be proteotoxic when used in Mycobacterial species, Sarah Fortune’s lab screened 11 other...

... labs and hinders experiments is mycoplasma contamination. Mycoplasma contamination has been shown to ...routinely test for mycoplasma contamination. There are several commercially available mycoplasma detection kits...larger microbes such as yeast, fungi, or bacteria, mycoplasma can be extremely hard to detect with levels reaching...cells per ml before the media becomes cloudy. Mycoplasma compete with host cells for nutrients and can...the cell line’s growth and behavior. Signs of a mycoplasma contamination can be subtle and may include reduced...positive controls. Routinely test your cell line for mycoplasma and discard it if the culture has become overly...References 1. Hay, R.J., Macy, M.L. & Chen, T.R. “Mycoplasma infection of cultured cells.” Nature 339 (1989...

...subunit S12 of 30S ribosome Confers resistance to streptomycin Tn10   Confers resistance to tetracycline ...galK rpsL (StrR) endA1 nupG fhuA::IS2 DB3.1 Streptomycin  HB101 derivative. For propagating plasmids ...lacY1 proA2 rpsL20(Smr) xyl5 Δleu mtl1 DH10B Streptomycin MC1061 derivative. General cloning and storage...lacZYA-argF)U169, hsdR17(rK- mK+), λ– HB101 Streptomycin Hybrid of E. coli K12 and E. coli B (mostly ...proAB+ lacIq lacZΔM15] hsdR17(rK-mK+) JM110  Streptomycin  For storing plasmids that should not be Dam...proAB+ lacIq lacZΔM15] hsdR17(rK-mK+)  MC1061 Streptomycin Parent of DH10B/TOP10 and derived strains, common...lac-proAB) mcrA Δ(mcrBC-hsdRMS-mrr) λ-  Stbl3  Streptomycin Derived from HB101. For cloning into and storage...

...methionine synthesis (5). The budding yeast, Saccharomyces. cerevisiae, makes a great reference organism... or not proteins from the fission yeast, Schizosaccharomyces pombe, have the same function as their counterparts...yeast for budding geneticists: A primer on the Saccharomyces cerevisiae model system. Genetics 197, 33-48...yeast for young geneticists: A primer on the Schizosaccharomyces pombe model system. Genetics 201, 402-423...Surdin-Kerjan, Y. Metabolism of sulfur amino acids in Saccharomyces cerevisiae. Microbiol Mol Biol Rev 61, 503-532.... et al. Functional characterization of the Saccharomyces cerevisiae genome by gene deletion and parallel...

...where recombination has occurred. Neomycin, puromycin, and hygromycin resistance genes are commonly used...insert a resistance gene. The vector contains a neomycin resistance gene (NeoR) flanked by homology arms...remove a resistance gene. In this construct, the neomycin resistance gene is floxed so that it can be removed...the sequence of GFP is cloned together with the Neomycin resistance (NeoR) gene between exon 1 and 3 of...

...from Kluyveromyces drosophilarum), R (from Zygosaccharomyces rouxii), B2 (from Zygosaccharomyces bailii...as low toxicity. FLP/FRT recombinases (from Saccharomyces cerevisiae) had been used successfully in Drosophila...bailii) and B3 (from Zygosaccharomyces bisporus.) Reporter constructs equivalent to site-STOP-site-myrRFP...

... ug/mL Bleomycin glycopeptide Induces DNA breaks Bactericidal 5-100 ug/mL Erythromycin macrolide...Class Mode of Action*   Working Concentration** Kanamycin aminoglycoside Binds 30S ribosomal subunit; ...mis-translation Bactericidal 50-100 ug/mL Spectinomycin aminoglycoside Binds 30S ribosomal subunit;...protein synthesis Bactericidal 7.5-50 ug/mL Streptomycin aminoglycoside Inhibits initiation of protein...

...result in a spectinomycin resistant phenotype in bacteria. While there are many known spectinomycin resistance...generate bacteria resistant to both spectinomycin and streptomycin, but could also be used for studying...five dispersed regions of the target gene of spectinomycin, rpsE. These results not only uncovered novel...

... gentamicin, hygromycin, kanamycin, nourseothricin (clonNat), spectinomycin, streptomycin, tetracycline...maintain a glycerol stock. Choose from: ampicillin, apramycin, blasticidin, bleocin (zeocin), chloramphenicol...

..., and is routinely tested for mycoplasma contamination using mycoplasma detection kits. Cell line is maintained... tested for mycoplasma contamination. To date, Addgene has never had a case of mycoplasma contamination...normalization purposes are being used. Quality Control Mycoplasma Our 293T cell line was obtained directly from...

...SpCas9 is fused to the puromycin resistance gene through a 2A peptide. The puromycin resistance gene enables...causes the ribosome to translate Cas9-2A and the Puromycin resistance gene as separate peptides. PX459 This...predecessor, which had a point mutation in the puromycin resistance gene that made it less effective in...

...80S; inhibits chain elongation 100-800 ug/mL Hygromycin B hygB HeLa, NIH3T3, CHO, Jurkat T cells Blocks...80S; inhibits chain elongation. 50-500 ug/mL Puromycin pac HeLa, 293HEK, Jurkat T cells Inhibits protein...do not confuse this with G418 (aka Geneticin). Neomycin should not be used for mammalian expression – ...confers G418 resistance; however, like gentamicin, neomycin is typically used to suppress bacterial growth...

...10.1093/genetics/iyae126.   Gene disruption in Mycobacterium abscessus using a dual-plasmid CRISPR/Cas9 system...dual-plasmid CRISPR/Cas9 system for gene disruption in Mycobacterium abscessus, an increasingly common human pathogen...-based systems have been developed for other mycobacteria, but such systems don’t always work across species...for rapid and efficient genetic disruption in Mycobacterium abscessus. J Bacteriol., 206(3), e0033523. https...