...mrr-hsdRMS-mcrBC) Phi80lacZDeltaM15 Delta-lacX74 recA1 araDelta139 D(ara-leu)7697 galU galK rpsL (StrR) endA1 nupG ...+B+ lacIq ∆(lacZ)M15 zzf::Tn10 (TetR) ∆(ara-leu) 7697 araD139 fhuA ∆lacX74 galK16 galE15 e14- Φ80dlacZ...mrr-hsdRMS-mcrBC) Phi80lacZM15 Delta-lacX74 recA1 araD139 Delta(ara-leu)7697 galU galK rpsL (StrR) endA1 nupG ...Invitrogen F– mcrB mrr hsdS20 (rB–, mB–) recA13 supE44 ara-14 galK2 lacY1 proA2 rpsL20 (StrR ) xyl-5 λ– leu ...indicated, the antibiotic powder can be dissolved in dH 2 0. Addgene recommends making 1000X stock solutions...million DNA fragments, which is far too many to separate from each other for the purposes of isolating ...recombinant DNA technology, which enables the preparation of more managable (i.e., smaller) DNA fragments...

...pHelper: 1 RepCap OR 3 Transfer Plasmid : 5 pHelper: 2 RepCap Select what type of tissue culture dish you...goes, “failing to plan is planning to fail.” Preparation is a key step to any experiment and can help ...

...Addgene swag. Good luck and happy naming!   Fig. 2: The mascot trio!     ...our new antibody mascot, who's joining us in preparation for our upcoming affinity reagent distribution...

...Halloween is only 2 days away and here at Addgene we couldn’t be more excited! As a small, tight-knit...(old board games and movies can provide great character costumes). All of this costume planning is for...

...expressed as a fusion with your favorite protein and (2) a synthetic fluorophore-contianing “ligand” that ...cell 58.4 (2015): 644-659. PubMed PMID: 26000849. 2. Grimm, Jonathan B., et al. "A general method to improve...: 26575238. 5. Crivat, Georgeta, and Justin W. Taraska. "Imaging proteins inside cells with fluorescent...

...connectivity to other data sources used in research; (2) connectivity to other tools used in research; and...by making it easy to get data out of the ELN; and (2) facilitate inter-operability with other research ... delve deeper and look specifically at the characteristics and capabilities an ELN needs to facilitate...

...blotting  In a western, proteins are: (1) separated by size, (2) transferred to a membrane, and  (3) detected...system-specific sandwiches.   Figure 2: The western blot dry apparatus includes a stack of filter paper, ...range to be separated on a single gel.  Immobilizing proteins on a membrane Once separated, the proteins... process. The sample, a mixture of protein is separated in a gel, transferred to a membrane, stained with...tertiary structures remain intact and the protein is separated through a matrix by charge. In a denaturing western...protein is denatured to its primary structure and separated by size with smaller molecules moving more quickly... we will be focusing on denaturing westerns.  Separating a protein mix by size Sodium dodecyl sulfate-...

...to a specific area at the plasma membrane (Figure 2). Just express the plasma membrane-localized iLID ...cells will be ready for the spotlight.    Figure 2: Opto-RhoGEFs to control Rho GTPase activity. A) Schematic...cytoskeleton. They provide cells with physical structure, separate chromosomes during mitosis, support transport ... called a katana. Opto-katanin consists of two separate blue light-sensitive systems, VVDfast to control...your protein of interest and express the other separately, then use blue light to generate CRY2-CIB1 oligomers...

...would label everything but the CD8+ T cells (Figure 2). Even better, we can use the same fluorophore for...gating on the negative population.     Figure 2: Dump gating labels all unwanted cell lineages with...conjugated to a biotin tag but not a fluorophore. In a separate step, cells are incubated with a fluorophore-conjugated...the intranuclear ones. You do not need to use a separate fix/perm cytosolic procedure in that case.   ...cell population. To detect non-surface markers, a separate intracellular labeling protocol is necessary, ...

...melanogaster." Molecular and cellular endocrinology 215.1-2 (2004): 1-10. PubMed PMID: 15026169. Soo, Rochelle...algae host for reproduction. Snail parasite Leucochloridium is a parasitic worm that infects a snail’s eyestalks...Brusca and Gilligan, 1983). Grasshopper parasite The parasitic worm Spinochordodes tellinii‘s larvae makes...ve also carved pumpkins (yes, Blugene made an appearance there too) and shared fascinatingly creepy science...Tongue-eating louse Cymothoa exigua is a marine parasite that enters fish through its gills. It attaches...replacement in a marine fish (Lutjanus guttatus) by a parasitic isopod (Crustacea: Isopoda)." Copeia 1983.3 (1983...

... a template, copying the base edit. Figure 2: Cytidine deamination takes place on the free strand...SpCas9 base editors with editing windows as small as 1-2 nucleotides. To reduce off-target effects associated...formation. To further decrease indel formation 1.5-2 fold, the team fused bacteriophage protein Gam to ...K., Arazoe, T., Yachie, N., Banno, S., Kakimoto, M., Tabata, M., Mochizuki, M., Miyabe, A., Araki, M.,...., Slaymaker, I. M., Yen, J., Young, L. E., & Ciaramella, G. (2020). Directed evolution of adenine base...Schwaegerle, N. D., Peraro, L., Young, L., Lee, S.-J., Ciaramella, G., & Gaudelli, N. M. (2023). Improved cytosine...., Hara, K. Y., Shimatani, Z., & Kondo, A. (2016). Targeted nucleotide editing using hybrid prokaryotic...

...environmental contaminants.  Figure 2: An Addgenie works in the tissue culture hood while wearing...doi.org/10.1290/1071-2690(2002)038<0086:caeomi>2.0.co;2 Uphoff CC, Drexler HG (2011) Detecting Mycoplasma ...of the causes of contamination. If possible, a separate lab should be made available for cell culturing...keeping them in the pockets of your lab coats. Have separate footwear for the lab to avoid bringing in environmental...wide range of mycoplasma. These methods cannot guarantee complete removal of mycoplasma but, can get rid...made permanent damage to the cell culture it is irreparable. Some cells may die due to longer incubation ...Microbiol .44: 42–49  Uphoff Cc, Drexler Hg (2002) Comparative Antibiotic Eradication of Mycoplasma Infections...

...either (1) a lack of gRNA and/or Cas9 expression or (2) a lack of efficient target cleavage in cells expressing...present in a single lentiviral transfer vector or separate transfer vectors May contain reporter gene (e....

...wheat germ, HeLa, and Leishmania (Gagoski, et al 2015[2]). The proteins encoded in this set range in size ...to generate Illumina sequencing libraries using a 2-step PCR-based protocol and are compatible with single-end...centered within GFP/RFP bicistronic reporters characterized in Cambray et al., 2013. These terminators can...size from 4 to 156 kDa, enabling the user to characterise the efficiency of their cell-free system in correlation...

...tagging." Nature methods (2015). PubMed PMID:26322837. 2. Stadler, Charlotte, et al. "Immunofluorescence and...the fusions had some negative effect on these parameters. IT5 and IT6 displayed good binding efficiency...

...Agrobacterium infection. At approximately 7 kb, pSiM24 is 2 and 6 kb smaller than commercially available pCAMBIA...duplicated enhancer domains and characterized in studies of tobacco, Arabidopsis, and corn. For both β-glucuronidase...plant infection, as well as plasmid yield during preparation in E. coli. Addgene depositor Indu Maiti has ...functions in multiple species, including tobacco and Arabidopsis. Improving Agrobacterium binary vectors Vectors...efficiently inherited in both transgenic tobacco and Arabidopsis. Finally, Sahoo et al. demonstrate that this ...

...elution choices: TE, Tris buffer, or water Figure 2: Tris-EDTA, Tris, or water? The buffer you choose ..., destabilizing the molecule and favoring the separation of the two helixes. Low pH instead results in... buffer for preserving the stability of your preparation for a long time. Tris buffer controls the pH,... if the priority is to directly use your DNA preparation in any downstream application involving enzymatic...buffer is that it allows better control of the pH preparation stabilizing the plasmid, and still allowing most...at -20°C or even at -80°C, in this case your preparation can be eluted in water or in your buffer of preference...

...blood are reprogrammed immediately after collection [2]. PBMCs display unique epigenetic signatures that ...PMID: 27882265. PubMed Central PMCID: PMC5118044. 2. Kim, Y., et al., The Generation of Human Induced ...vectors encoding reprogramming factors behave as separate extrachromosomal elements in the nucleus of the...

...sequence is unique compared to the rest of the genome.  2) The target is present immediately upstream of a Protospacer...utilized allowing for PAM flexibility. Fig. 2: Prime editor engaging target DNA. The reverse transcriptase...Walton et al., 2020). Further, this group also characterized a second Cas9 variant, SpRY, which cuts at NRN...

...faster when using a dual guide system.   Fig. 2: Size screening for large deletion events.   ... Biol 17: https://doi.org/10.1186/s13059-016-1012-2 Find more CRISPR Resources at Addgene: Read our CRISPRGuide...visualized by running your PCR on a gel. It’s still paramount that you sequence clones to verify that the deletions...