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We narrowed to 168 results for: plasmid for e coli

Showing: 121 - 150 of 168 results
  1. Anatomy of a Plasmid Page at Addgene

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    Blog Post
    ... best bet is the Plasmid Map: We auto-generate a plasmid map based off of the plasmid sequence and feature...name of the strain.  Most Addgene plasmids are maintained in the E. coli lab strain DH5a, but others may...colleagues. Top of the plasmid page The ‘breadcrumb’ trail at the top of any plasmid page gives the hierarchy... to the individual plasmid page. The depositing lab page contains all the plasmids from the same lab while...contains all the plasmids deposited from the same publication. Next up, we have the plasmid name: This is...other plasmids with similar names in the collection, but you can use this name to find this plasmid from...A quicker way to find this plasmid through search is... The Addgene plasmid number: This is the unique...
  2. Pooled Library Amplifications

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    Blog Post
    ...recombined plasmid to overtake full-size plasmids (or for smaller plasmids to overtake larger plasmids, if the...How many unique plasmids does your library contain?), the variance of size of plasmids in your library...transformation amplification protocol is the Chong Zhang E. coli Genome-wide Inhibition Library. DNA and cell amounts...lead to issues such as recombination and loss of plasmid diversity, so Addgene strives to distribute aliquots...library (How important is it that every single plasmid in your library is present, and how will you ensure...30 to 200 times as many colonies as individual plasmids in your library. As an example, if you want 100x... your library, and your library contains 5,000 plasmids, your goal for your transformation would be to...
  3. Antibodies 101: Affinity Reagents

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    Blog Post
    ...specificity and binding affinity, can be produced in E. coli, and are both soluble and easily engineered.  Monobodies...Resources Additional resources on the Addgene blog Plasmids 101: Aptamer FluorophoresAntibodies 101: scFVs...available from AddgeneBrowse our recombinant antibody plasmid collection References Grumezescu, A. M. (2018)....
  4. CRISPR 101: Ribonucleoprotein (RNP) Delivery

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    Blog Post
    ...express and purify His-tagged Cas9 from E. coli using a plasmid such as pET-28b-Cas9-His from Alex Schier...commonly performed by transfecting cells with a plasmid, such as PX459, which encodes Cas9 and contains...chosen for both Cas9 and gRNA expression       Plasmid DNA may be incorporated into the genome     Off-target...targets with similar efficiency as compared to plasmid-based expression of Cas9/gRNA and can be used for... as CMV or EF1A promoters found in many CRISPR plasmids) are not expressed. Because this method does ...fluorophore tagged dCas9 may require the use of plasmid or viral-mediated delivery. You’ve decided to...experimental systems that may not be amenable to plasmid based methods. One of the most common techniques...
  5. Hot Plasmids - August 2020

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    Blog Post
    ...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...They generated plasmids tagged with EB3 to mark the growing tips of microtubuli and plasmids tagged with ...and dCas9-VPR targeting plasmids can be found at Addgene. Check out these plasmids for piggyBac gRNA delivery...highlights from recent CRISPR plasmids. To find all of the CRISPR plasmids available from Addgene, head... Find the plasmids at Addgene The MoClo CRISPR/Cas Toolkit for plants includes 95 plasmids consisting...Nanobodies are easily expressed in the cytosol of E. coli, however the nanobody proteins tend to get stuck...
  6. Plasmids 101: Choosing an Antibiotic Resistance Gene

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    Blog Post
    ...resistance DNA plasmids are traditionally grown in bacteria as a way to produce more of the plasmid, isolate...isolate individual plasmids, or express a gene contained in a plasmid. During the propagation and/or transformation...receive a copy of the plasmid or will lose it during division. These non-plasmid-containing bacteria can...Over-Agar Antibiotic Plating Plasmids 101 eBook Resources on the Addgene blog Plasmids 101: Antibiotic Resistance...Resistance Plasmids 101: Blue-White Screening Choosing Your Perfect Plasmid Backbone  ...Plasmids need antibiotic resistance genes so that you can specifically isolate bacteria containing your...your experiment — to grow, express, or isolate plasmids. The presence of an antibiotic resistance gene...
  7. CRISPR Antimicrobials

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    Blog Post
    ...resistance genes in E. coli using plasmid and phagemid delivery systems. Phagemids are plasmids containing a ...genome is lethal. If this DSB occurs on a plasmid, the plasmid will be eliminated from the bacterium, which...sequence-specific, antimicrobial, plasmid-based, CRISPR systems (Find the Citorik Plasmids here). In both cases, ...sequence results in cell death. Plasmid targeting may result in plasmid loss and antibiotic resensitization...this resistance gene being encoded by a high-copy plasmid. In the case of the genomically encoded quinolone...RNAs (crRNAs) targeting either genomic (blue) or plasmid-based sequences (red). Treating a mixed population...specifically targets bacteria with the given genomic or plasmid sequence from a mixed population without affecting...
  8. Viral Production at Addgene

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    Blog Post
    ...the transfer plasmid, a plasmid encoding rep and serotype-specific cap genes, and a plasmid encoding adenoviral...coli. In addition, plasmids are typically prepared using endotoxin-free plasmid purification kits.   ... all plasmids used for viral production are propagated in the endA-mutated NEB Stable strain of E. coli...contamination is minimized by using an endotoxin-free plasmid purification protocol. To minimize the immunogenic...offline and discarded. Confirmation of the transfer plasmid Addgene uses a rigorous barcode matching system...undergoes PCR with primers targeting the transfer plasmid used in the transfection. PCR products are analyzed... system and PCR-based confirmation of transfer plasmids used in Addgene’s viral production. Barcoding ...
  9. Plasmids 101: Secondary Nanobody Toolbox

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    Blog Post
    ...Activity Learn more about protein tags Read about E. coli strains for protein expression Additional Resources... How to Use the Nanobody Toolbox Find Nanobody Plasmids Structural comparison As the name suggests, ... Species Antibody Subclass Nanobody Addgene Plasmid Rabbit N/A* TP897 pTP1183 (1x cysteine) pTP955...Nanobody In Bacteria: Table 2 also has a link to the plasmid you’ll need to express your nanobody of choice ...column. bdNEDP1 protease can be generated using plasmid pDG02583. Label the Nanobodies: Purified nanobodies...Journal of Cell Biology. PubMed PMID: 29263082 Find plasmids from this publication at Addgene. 2. Bates, M... on Addgene.org Find more nanobody expression plasmids here Additional Resources Looking for monoclonal...
  10. Plasmids 101: The Promoter Region – Let's Go!

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    Blog Post
    ...trp High levels of gene expression Promoter from E. coli tryptophan operon Repressible Gets turned off with...Once you've gotten to know what a plasmid is in general, it's time to take a look at some of its parts...origin of replication are necessary to replicate plasmids and make sure the cells maintains it. Here, we'll...the promoter, which is necessary for getting the plasmid express our gene of interest. This element is responsible.... This means that the promoter present in your plasmid backbone must to be compatible with the type of...transcript, you will also need to make sure your plasmid has a promoter suited to working in your host organism...development-specific promoters available to scientists. Plasmids are oftentimes put to therapeutic uses, and in...
  11. Cpf1: A New Tool for CRISPR Genome Editing

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    ...Francisella Cpf1 (FnCpf1), they employed an E. coli plasmid depletion assay to discover FnCpf1’s PAM sequence...system." Cell (2015). PubMed PMID: 26422227. Find plasmids from this publication at Addgene. 2. Zetsche,....1 (2016): 31-34. PubMed PMID: 27918548. Find plasmids from this publication at Addgene. 3. Kim, Daesik...Biotechnol. (2016). PubMed PMID: 27272384 Find plasmids from this publication at Addgene. Kleinstiver...Biotechnol. (2016). PubMed PMID: 27347757. Find plasmids from this publication at Addgene. Makarova, Kira...PMID: 26411297. Resources at Addgene Find CRISPR plasmids at Addgene Read Addgene's CRISPR Resource Guide...
  12. Tips for Screening with Yeast Two Hybrid Systems

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    Blog Post
    ...transforming both fusion plasmids, or mating two single haploid yeast fusion plasmid transformants into a ...reporter system (e.g. the Gateway-compatible bait Y2H plasmid pEZY202  from Yu-Zhu Zhang’s lab) is an example...fusions for analyzing gene expression in Escherichia coli and yeast. Methods Enzymol. 1983;100:293-308. PubMed...Yeast Expression Vectors Find Bacterial Two Hybrid Plasmids Read our Blog Post About Yeast Vectors Read our... Blaudez, D., Sanders, D., Revuelta, J.L., Boles, E., André, D., and Frommer, W.B. K+ channel interactions...
  13. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

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    ...analyzing the plasmid and viral DNA sequences and expression profile at multiple steps from plasmid amplification...recombination persists even in recombinase deficient E. coli and under conditions used to minimize recombination...to flip back into the inverse orientation (See Plasmids 101: FLEx Vectors). While this strategy greatly...recombination that are typically employed during viral plasmid amplification (Fischer, KB. 2019). This means that...of this finding, we now include guidance on the plasmid page of DIO/FLEx AAV vectors where this occurs ... mechanism of spontaneous reversion during AAV plasmid amplification. On the left homologous pairs of ....2020.618020 Additional resources on the Addgene blog Plasmids 101: FLEx Vectors Important Considerations When...
  14. Site Directed Mutagenesis by PCR

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    Blog Post
    ...mutagenesis of Plasmids. A) Generation of mutagenized plasmids. PCR primers (green) amplify the plasmid template...with the nuclease-resistant nicked plasmid (the PCR product). Plasmids are isolated from the resulting colonies...the final circular plasmid product pictured here. B) Screening of recovered plasmids for the mutation of...Smaller plasmids (~3 kb) are generally more efficiently amplified than larger constructs, but plasmids as ...DpnI) to eliminate the parent plasmid from the PCR products, the plasmid template should be isolated from...have taken up the mutagenized plasmid. Note that any residual parental plasmid (usually from incomplete DpnI...additional restriction site into your plasmid, then, when the plasmid is digested with the appropriate restriction...
  15. Tips and Tricks for Using Golden Gate Modular Cloning (MoClo)

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    Blog Post
    ...blog Plasmids 101: Modular Cloning Plasmids 101: Modular Cloning Applications and Kits Plasmids 101: ...through your plasmid stocks. To avoid cross-contamination, we recommend retrieving plasmids from your original...Calculation for the plasmid volume for MoClo experiments. The term ‘insert’ refers to plasmids carrying each...after each transformation and/or use different E. coli strains with higher transformation efficiency. ...will find some useful tips to help you make your plasmid construction with MoClo a walk in the park.  Before...promoter, a coding sequence, a terminator, or other plasmid part? Do you want to fuse your protein to any tags...restriction enzymes, and it will create the resulting plasmid map and identify any mismatches that are preventing...
  16. 28 Hot Plasmid Technologies from 2015

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    Blog Post
    ...new origin of replication increases plasmid copy number in E. coli. The authors used pXPG to study the...amazing plasmid technologies developed by our community of depositors. With over 40,000 plasmids avaliable...amazing new plasmid tools that have come through our doors in 2015. Do you have a favorite new plasmid tool ...system plasmids are available through Addgene and are already very popular. Use these plasmids to increase... can visualize: Cell Structure Plasmids: Cell Structure Plasmids: ER mCherry-Sec61 beta BFP-Sec61...luciferase reporter gene plasmid pGL3 pGL3 luciferase reporter gene plasmids have been used extensively...find all of our Hot Plasmid articles on our website. We update the our Hot Plasmids articles every quarter...
  17. Plasmids 101: Biotinylation

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    ...PMC4884002. 10. Boer, E. de, Rodriguez, P., Bonte, E., Krijgsveld, J., Katsantoni, E., Heck, A., … Strouboulis...biotinylating proteins with BirA. You can find plasmids that contain both AviTag and BirA at Addgene! ...sequestering biotin from being absorbed. In 1940 Esmond E. Snell was able to purify the protein responsible ...). References 1. Chapman-Smith, A., & Cronan, J. E. “Molecular Biology of Biotin Attachment to Proteins...PMID: 10064313. 2. Chapman-Smith, A., & Cronan, J. E. “The enzymatic biotinylation of proteins: A post-.... and R. L. H. “The Discovery of Avidin by Esmond E. Snell.” J Biol Chem 279.e5 (2004). PubMed PMID: 20353954...of the E.coli BPL, BirA. If you recall, BirA biotinylates only one lysine in the entire E.coli proteome...
  18. CRISPR History and Development for Genome Engineering

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    ...posts , or find CRISPR plasmids for your research. Addgene has CRISPR plasmids for the following models...Repeat) sequences were initially discovered in the E. coli genome in 1987, but their function as a safeguard...Mammalian systems (Human, mouse, and rat) Bacteria ( E. coli , Streptococcus , Streptomyces , and others) Drosophila... in the field, and enabling access to multiple plasmids used for various platform applications. For more...alkaline phosphatase isozyme conversion in Escherichia coli, and identification of the gene product. J Bacteriol...Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E. 2012. A programmable dual-RNA-guided DNA endonuclease... Saunders SJ, Barrangou R, Brouns SJ, Charpentier E, Haft DH, Horvath P, Moineau S, Mojica FJ, Terns RM...
  19. Recombinase-based State Machines Enable Order-dependent Logic in vivo

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    ...implement RSMs in E. coli with two plasmids (Figure 3 below): Input Plasmid: The input plasmid (high copy, ...inducible promoter (PPhlF).  Output Plasmid: The output plasmid (single copy, bacterial artificial chromosome...through Addgene. Their plasmid maps and GenBank files can be found on their plasmid pages. All two-input...indicated figures in our paper (1)) Input Plasmid Output Plasmid Fig. 3A pNR64 pNR160 Fig. 4A pNR220...two-input RSMs and three three-input RSMs. The plasmids used to construct these RSMs are listed in the...paper (1). These parts are also annotated on the plasmid maps and GenBank files uploaded to Addgene. We ... Visit Our SynBio Pages Check Out More Lu Lab Plasmids Find More Genome Engineering Tools  ...
  20. Adenovirus Plasmids

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    ... Viral Plasmids Adenoviral Plasmids Adenoviral Plasmids Shuttle Plasmids Adenoviral Plasmids Adenoviral...16398 BJ5183 E. coli strain for recombination between shuttle plasmid and adenoviral plasmid Vogelstein ... generally need to use two plasmids: A shuttle plasmid (or transfer plasmid) containing the transgene ...for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail help...Browse shuttle plasmids, adenoviral backbone plasmids, and bacterial strains for adenoviral vector production...for recombination with the adenoviral plasmid. An adenoviral plasmid containing the rest of the adenoviral...with the shuttle plasmid. This collection provides a curated selection of popular plasmids that can be used...
  21. Antibodies 101: Introduction to Antibodies

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    Blog Post
    ... be used for. Addgene’s Antibody Plasmid Collection includes plasmids that express antibodies, nanobodies...Engineering production of functional scFv antibody in E. coli by co-expressing the molecule chaperone Skp. Front...immunoglobulin genes, recombinant antibodies are plasmid-based. Their sequences can be optimized for specificity...protein (Wang et al., 2013). These proteins are also plasmid-derived. However, a disadvantage to ScFv is their...nanobodies, or synbodies, can be produced from plasmids, saving time and money. Nanobodies and synbodies...expression in cells that have been transfected with a plasmid encoding GFP. Actin, a common internal control ... as well as tools for developing and producing plasmid-based antibodies - and we’re always growing our...
  22. CRISPR Plasmids - Base Edit

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    ...for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail help...Bacteria ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Return to top Plant ID Plasmid Gene...selection of CRISPR plasmids and resources. Find more CRISPR functions along with plasmids categorized by ...CRISPR plasmids for precisely editing DNA bases without inducing a DNA break. CRISPR...CRISPR Plasmids: Base Edit Mammalian Bacteria Plant Yeast Zebrafish Resources Two classes of base editors ...created by directed evolution of the Escherichia coli TadA, a tRNA adenine deaminase. Like cytosine base...search the tables below for CRISPR base editing plasmids. To learn more about base editing and other CRISPR...
  23. 15 Hot Plasmids from 2017

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    Blog Post
    ...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These brief articles...articles provide a synopsis of a plasmid or group of plasmids' functions and applications. We hope that... you to find and use the plasmids you need. You can find all the hot plasmids from 2017 below. With over...over 50,000 plasmids, we can't write posts for every great plasmid that comes into the repository, but...write about your plasmids in a future blog post. No time to read? Listen to our hot plasmids segment on the...editing for precise genome modification. Plasmid ID Plasmid Name Cas9 Variant Speacial Features 85169...Bacterial Expression Construct SpCas9 Inhibition (E. coli) Mammalian Expression Construct SpCas9 Inhibition...
  24. Empty Backbones - Choosing Your Perfect Plasmid Backbone

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    ...for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail help... Plasmid Collections Empty Backbones Choosing Your Perfect Plasmid Backbone Empty backbones...Collection NLS Nuclear localization dCas9 plasmid - dCas9 expression plasmid without effector fusions; 3X Flag... of plasmids for packaging and expression, see our dedicated Adeno-associated Virus (AAV) Plasmids page...collection of plasmids for packaging and expression, see our dedicated Lentiviral Plasmids page Retroviral...contains AdEasy®-1 plasmid View our Adenovirus Plasmids page for more information Genome...of your cells will take up your plasmid. Thus, many plasmids have markers on them so that you...
  25. Promega Plasmid Collection

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    ...Promega Plasmid Collection Promega Plasmid Collection Ordering & Availability All Plasmids Plasmid Technology...our help center . Plasmid Collection Browse and sort the table below to find plasmids in the collection...a new window) on Promega.com. ID Plasmid Description Industry Plasmid Technology Overview This collection...Browse Addgene's Luciferase Plasmid Collection Explore other Promega plasmids available at Addgene Find ...for various protein expression systems including E. coli , mammalian cells, and cell-free systems. NanoBiT...operational complexity. This unique collection contains plasmids developed by Promega scientists and designed to...HaloTag, SmBiT, and LgBiT. Ordering & Availability Plasmids from the Promega collection are available in two...
  26. COVID-19 Resources

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    ... Plasmid Collections COVID-19 – Plasmids and Resources COVID-19 and Coronavirus Plasmids & Resources...Antibodies SARS-CoV-2 Plasmids Other Coronavirus Plasmids Mammalian Targets General Plasmid Tools Pooled Libraries...Brian Geiss Plasmids SARS-CoV-2 Plasmids Browse the tables below to find popular plasmids expressing genes...to top Other Coronavirus Plasmids: SARS-CoV, MERS, BAT-CoV Plasmids ID Plasmid Description Industry PI ...Ginkgo Bioworks to distribute SARS-CoV-2 plasmids for E. coli , yeast, and mammalian expression, and spike...for other plasmids that should be added to this list? Fill out our Suggest a Plasmid form or e-mail help...for-profit entity, please see our Plasmid Distribution to Industry page. ID Plasmid Description Industry PI Additional...
  27. Viral Production

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    Collection
    ...the transfer plasmid, a plasmid encoding Rep and serotype-specific Cap genes, and a plasmid encoding adenoviral...coli . In addition, plasmids are typically prepared using endotoxin-free plasmid purification kits. Resources...all plasmids used for viral production are propagated in the endA -mutated NEB Stable strain of E. coli...contamination is minimized by using an endotoxin-free plasmid purification protocol. To minimize the immunogenic...off-line and discarded. Confirmation of Transfer Plasmid Addgene uses a rigorous barcode matching system...experiment using our virus protocols . Browse our plasmid collection of popular lentiviral vectors ....
  28. CRISPR Guide

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    ...36593413 Vo, P. L. H., Ronda, C., Klompe, S. E., Chen, E. E., Acree, C., Wang, H. H., & Sternberg, S. H... R., Hanna, R. E., Hegde, M., Donovan, K. F., Strand, C., Sullender, M. E., Vaimberg, E. W., Goodale, ... the DNA and replaces it with another base. In E. coli , this strategy yields C to A base edits, whereas...harnessed for antiviral mechanisms in bacteria ( E. coli ). Like the other Cas enzymes, the Cascade-Cas3...edits. PASSIGE ( P rime- E diting- A ssisted S ite- S pecific I ntegrase G ene E diting), developed by the.... PMID: 26735016 Lee, J. K., Jeong, E., Lee, J., Jung, M., Shin, E., Kim, Y., Lee, K., Jung, I., Kim, ..., K. M., Aach, J., Guell, M., DiCarlo, J. E., Norville, J. E., & Church, G. M. (2013). RNA-Guided Human...
  29. CRISPR References and Information

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    ...Addgene CRISPR Resources CRISPR Plasmids Addgene's collection of CRISPR plasmids. CRISPR Guide Addgene's guide...and activation). Genome support for bacteria ( E. coli, B. subtilis ), yeast ( S. cerevisiae ), worm (...guide to using CRISPR plasmids in your lab, from knockouts to pooled library screening. CRISPR eBook Addgene's...barcoding. CRISPR Kits Addgene's collection of CRISPR plasmid kits. CRISPR Software Sanger Indel Analysis ICE... Feng Zhang Lab . Protocols Lab(s) Description Plasmids in protocol Download protocol Addgene CRISPR pooled...gRNA design and cloning for Cas9 orthologs Cas9 plasmids PDF, 107 KB Chen and Wente Zebrafish: gRNA cloning... (Cre recombinase) PDF, 105 KB Jaenisch CRISPR plasmid summary for transcriptional activation dCas9 activators...
  30. Hot Plasmids September 2018 - Optogenetics, RNA Localization, Fluorescent Protein, and Base-editing Tools

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    ...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles ...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...about a recent plasmid deposit please sign up here.   Listen this episode of Hot Plasmids! Light controls... with broccoli aptamers or MS2 tags using the same number of fluorophores. You can find plasmids containing...Hahn's lab at the University of North Carolina (plasmids at Addgene). It consists of two domains, LOV2 ...the Reuben Harris lab has deposited a reporter plasmid that allows for rapid fluorescence-based quantification...editing complexes are more efficient than the BE3 plasmid containing rat APOBEC1. Finally, they show that...
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