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We narrowed to 131 results for: transformation

Showing: 21 - 40 of 131 results
  1. Tips and Tricks for Using Golden Gate Modular Cloning (MoClo)

    Type
    Blog Post
    Published
    July 8, 2025, 1:15 p.m.
    ... after each transformation and/or use different E. coli strains with higher transformation efficiency....colonies) after each consecutive assembly and transformation round, due to the decrease in ligation efficiency...’re cloning in high-throughput, we recommend transforming into chemically competent cells in a 96-well...have what look like positive colonies on your transformation plates. However, even though the MoClo system... always screen multiple colonies after each transformation. This could be as few as 1–2 colonies if you...
  2. Cultivating Community Science at BosLab

    Type
    Blog Post
    Published
    Nov. 2, 2017, 1:35 p.m.
    ...December 2015 we held our first intro to bacterial transformation course (read our blog post on it and to see...consisting of over twenty members and volunteers. Transforming citizens into scientists Anyone can become a...dedicated time at BosLab, he became our bacterial transformation guru and instructor for our intro to biohacking...Addgene and are eager to do our first yeast transformations! Dennis Oleksyuk, Raphael Laurenceau, and Tami...
  3. Troubleshooting Your Plasmid Cloning Experiment

    Type
    Blog Post
    Published
    Sept. 24, 2019, 12:55 p.m.
    ...the specific reaction you are running. Transformation Transform a few microliters of your ligation reaction...The PCR could fail to produce a product, the transformation may not result in any cells, or all the colonies...fragments If you don’t get any positive clone, try transforming the ligation mix in different bacterial strains...
  4. Technique: Probe Phage Genomes for Host Binding Proteins

    Type
    Blog Post
    Published
    Sept. 29, 2016, 2:30 p.m.
    ...certain modifications to DNA bases.  Ligate and transform your resultant plasmid libraries into chemically...chemically competent E. coli and analyze transformants for appropriately sized inserts. We consider a library...culture, dilute and plate the complete library of transformants for single colonies (this usually requires 10...10-20 plates) (see box below). This initial transformation should be performed using a highly competent...extract plasmid DNA directly from the colonies. Transform the resultant plasmid DNA into E. coli BL21 (DE3... protein production), then dilute and plate transformants for isolated colonies (150-200 per plate) across...
  5. pSiM24: Simplifying Plant Genetic Engineering

    Type
    Blog Post
    Published
    Sept. 29, 2015, 2:30 p.m.
    ...binary vectors are quite large (>10 kb.) Since transformation efficiency is inversely proportional to size...ORI, they achieved much higher DNA yield and transformation efficiency in both types of bacteria. They ... promoter, and it can be used to select both transformed Agrobacterium and infected plants. Further opportunities...
  6. Plasmids 101: The Wide World of Natural Plasmids

    Type
    Blog Post
    Published
    Oct. 8, 2024, 1:15 p.m.
    ...Horizontal gene transfer can occur naturally through transformation (cellular uptake from the environment), transduction...one popular method scientists use to promote transformation, but it isn’t just for lab specimens — there...plasmid are now used routinely in biotechnology to transform plant cells. Degradative plasmids: microbial hardiness...Plasmids 101: What is a Plasmid? Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection...
  7. CRISPR-mediated Plant Base Editors

    Type
    Blog Post
    Published
    Jan. 3, 2019, 1:35 p.m.
    ...72 hours post-transformation/transfection in a transient assay. For stable transformation, regenerated ...deaminases (ecTadA), fused it with dCas9, and transformed it into E. coli harboring the defective CamR ...genome edited plants via Agrobacterium-mediated transformation. We also constructed smaller vectors suitable...suitable for transient assays using protoplast transformation. Cytosine base editors for plant There are several...
  8. Plasmids 101: Origin of Replication

    Type
    Blog Post
    Published
    Nov. 11, 2020, 2:23 p.m.
    ...plasmids from the same group should not be co-transformed, so if you need two plasmids for an experiment... same incompatibility group should not be co-transformed. +Actual copy number varies. See below for ...stocks, agar stabs, or liquid cultures. Freshly transformed cells also give a higher plasmid yield that colonies...
  9. Viral Vectors 101: Preparing Pooled Libraries

    Type
    Blog Post
    Published
    July 13, 2023, 1:15 p.m.
    ... the pooled library is transformed into E. coli for amplification. Transformed E. coli are allowed to ...library. This is similar to amplifying a plasmid: transform E. coli with the pooled library DNA, allow the...red.    Unlike a standard single plasmid transformation, though, a pooled library has hundreds to millions...
  10. Pushing the Limits of DNA Assembly

    Type
    Blog Post
    Published
    Sept. 6, 2017, 1:01 p.m.
    ...target region. Assembled plasmids can be directly transformed into E.coli for plating, screening and generation..., generating expression cassettes for yeast transformation can take 5 days. NEBuilder shortens that time...inserted into a linearized cassette, ready for transformation into yeast that same day.  The field of synthetic...covalently joined, scarless DNA – perfect for transforma­­­tion or as a PCR template. Some of the applications...
  11. Plasmids 101: Blue-white Screening

    Type
    Blog Post
    Published
    June 4, 2015, 1:03 p.m.
    ...produces pigment from x-gal (included in the transformation plate medium), turning the bacterial colony...for blue-white screening Use a good control: Transform the backbone plasmid without insert. All colonies...components, and are rescued by genes supplied on the transformed plasmid. Such plasmids may contain genes that...
  12. 3 Challenges in Plant Synthetic Biology

    Type
    Blog Post
    Published
    July 22, 2014, 5:46 p.m.
    ... still in its infancy. A long timeline from transformation to testing and a lack of well-characterized...s made even more difficult by the fact that transformation methods have also been patented. In order to.... We will also be working with open source, transformation technologies like those pioneered by Ewen Mullins...
  13. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    Published
    Aug. 2, 2016, 2:30 p.m.
    ...methylation-dependent endonuclease (i.e. DpnI), and bacteria are transformed with the nuclease-resistant nicked plasmid (the...the adenosine of the GATC recognition site. Transformation: After the PCR reaction, no ligation is required...required since the E. coli you transform your PCR products into will efficiently patch up the DNA. The ...parental plasmid provides a mean for selecting for transformants which have taken up the mutagenized plasmid....
  14. CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9

    Type
    Blog Post
    Published
    Oct. 11, 2016, 2:30 p.m.
    ... More information on Agrobacterium-mediated transformation can be found in this blog post. The pDGE Dicot...Simplified schematic of agrobacterium-mediated transformation in plant cells. Agrobacterium tumefaciens can...plant genome. B) The Ti plasmid and helper are transformed into agrobacterium tumefaciens and exposed to...Gelvin SB (2003) Agrobacterium-Mediated Plant Transformation: the Biology behind the “Gene-Jockeying” Tool...
  15. Six Spooky Science Stories and Halloween at Addgene

    Type
    Blog Post
    Published
    Oct. 31, 2019, 1:06 p.m.
    ... the snail, the larvae reach the eyestalk and transform them into a pulsating display imitating a moving...grasshoppers and crickets. When it’s ready to transform into an aquatic adult, it forces its insect host...
  16. Plasmids 101: Positive and Negative Selection for Plasmid Cloning

    Type
    Blog Post
    Published
    Aug. 22, 2019, 12:43 p.m.
    ...includes an antibiotic resistance marker. After transformation, plate the cells on medium containing the corresponding...selection will select for cells that have been transformed with the plasmid backbone (with or without your...
  17. Tag Your Favorite Yeast Genes with Ease

    Type
    Blog Post
    Published
    Nov. 19, 2013, 2:37 p.m.
    ...product is introduced into the cell by direct transformation. A given insert will typically contain both...selectable gene product for isolation of successful transformants. Addgene distributes several ready-to-use, modular...
Showing: 21 - 40 of 131 results