Skip to main content
Addgene
Open Menu

Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

Information

In Category

We narrowed to 629 results for: EMB

Showing: 61 - 80 of 629 results

  1. General Transfection

    Type
    Protocol
    ...drifted. Filter the solution through a 0.22 µm membrane. Aliquot 500–1000 µL into sterile tubes. Store...limited effect on cell growth. Last reviewed on: November 7, 2023...

  2. Protocol - Bacterial Transformation

    Type
    Protocol
    ... still get enough colonies for your next step. Remember that each of these shortcuts will reduce the efficiency...field is applied to the cell/DNA mixture to induce membrane permeability. To do this you will need to have...

  3. Synthetic Biology - Networks and Gene Regulation

    Type
    Collection
    ...SynBio Resources SynBio Home Browse All SynBio Assembly Standards SynBio Blog Genome Engineering Cloning...ocurring and synthetic regulatory elements, pre-assembled genetic circuits, and higher level gene networks...

  4. Using a Light Microscope Protocol

    Type
    Protocol
    ...started in the lab! Introduction Microscopes are emblematic of biological research and are found in many ...focus. Pro-Tip As you increase your magnification, remember that the objectives are larger. Adjust your focus...

  5. Pipetting Protocol

    Type
    Protocol
    ...pipette tip and using the pipette. Last Update: September 2022 Video Watch the video for tips on pipetting...preloaded into tip boxes. In either case, once assembled into their boxes, pipette tips can be sterilized...

  7. Antibody Production

    Type
    Collection
    ...encoded by mammalian expression vectors, human embryonic kidney cells are transfected with the plasmid(...cells, separated by SDS-PAGE, transferred to a membrane, and labeled with the antibody and a species-specific...

  8. New England Biolabs Cell-Imaging Plasmid Collection

    Type
    Collection
    ...with a recognizable sub-cellular localization (membrane, nucleus, etc.) are also available Non-toxic to...pSNAP-CaaX Control Plasmid SNAP-tag Control for endomembrane (CaaX box) localization 101129 pSNAPf-Cox8A ...

  9. Allen Institute for Cell Science Plasmid Collection

    Type
    Collection
    ...101784 SLC25A17-mEGFP AICSDP-27 mEGFP Peroxisomal membrane protein PMP34 Peroxisomes 101785 TUBA1B-mTagRFP-T...AICSDP-42 mTagRFPT CAAX domain of K-Ras Plasma Membrane 114403 LMNB1-mTagRFP-T AICSDP-29 mTagRFPT Lamin...

  10. Neurodegeneration Research Collection

    Type
    Collection
    ...Neurodegeneration Research Collection Addgene has assembled the following to help you find information and...cases having at least one other affected family member. These familial cases, usually inherited in an ...

  11. Empty Backbones - Choosing Your Perfect Plasmid Backbone

    Type
    Collection
    ...commonly added to aid in protein purification. Just remember to remove the stop codon for C-terminal tags and...yeast expression vector that adds a SV40 NLS Myr Membrane localization pWZL-Neo-Myr-Flag-DEST - ...

  12. Plan Your Experiment

    Type
    Collection
    ...read our three tips to improve HDR efficiency . Remember that HDR also requires donor DNA to serve as template...can transfect at high efficiency, such as human embryonic kidney 293 (HEK293) cells. For more difficult ...

  13. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...Vector Addgene Plasmid 10878. Protocol Version 1.0. December 2006. Copyright Addgene 2006, All Rights Reserved... . 1996 272:263-267. PubMed . Asymmetry in the assembly of the RNAi enzyme complex. . Schwarz DS et. al...

  14. Troubleshooting and Optimizing a Western Blot

    Type
    Blog Post
    ... your membrane, run a double-membrane transfer with a psi membrane stacked behind the p membrane. If your...through the p membrane into the psi membrane, that’s a good indication that the p membrane pores are too...marked on the membrane (1) using the gel as a guide. The gel is then removed from the membrane (2) and the...troubleshoot.   Before you start… Mark your membrane! Membranes look the same from both sides, so it’s quite...appropriately sensitive. If using a PVDF membrane, you can dry the membrane, wet it with 20% methanol, and place...completely through the membrane or larger proteins not transferring completely onto the membrane. If you’re not...completely through your membrane to the other side), you can check by using two membranes during your transfer...

  15. Antibodies 101: Stripping and Reprobing Western Blots

    Type
    Blog Post
    ... your gel and membrane transfer as normal. Immediately after transfer, dry the membrane to maximize protein...manufacturer’s instructions for the membrane you are using. Dry nitrocellulose membranes are brittle, so proceed ... as normal! Stripping your membrane Alright, so you’ve dried your membrane and returned it to buffer, ...depend on your specific experiment. Remember, stripping your membrane will remove some of your sample —and...’ll want to: Rinse the membrane in water or fresh buffer. Cover the membrane in stripping solution and...a fume hood. Rinse the membrane in water or fresh buffer. Cover the membrane in stripping solution and...you need to start a whole new blot? Never fear: membrane stripping is here! Rather than start over, you...

  16. Human Germline Editing Using CRISPR

    Type
    Blog Post
    ...off-target editing sites in the embryos, as well as whole exome sequencing of embryo derived-ES cells.They also...editing between iPS cells and embryos make a case for conducting research in embryos should our society decide...sequencing to a small percentage of edited embryos. Since the embryos produced were not being used to establish...-free embryos to only 60%? Unfortunately, given the differences between CRISPR repair in embryos and iPS...this paper. Any hint of CRISPR editing in human embryos has been met with a storm of media coverage. But... replication. Indeed, they observed that 41/42 embryos were non-mosaic at 3 days post fertilization. Since...MYBPC3 allele. Interestingly, they found that the embryos underwent repair not from the single-stranded oligonucleotide...

  17. Technique: Probe Phage Genomes for Host Binding Proteins

    Type
    Blog Post
    ...nitrocellulose membranes and exposed to IPTG in order to induce protein expression. Cells on the membranes will ...orientation of each membrane on its plate before moving on. Then peel each membrane off the plate (saving...high salt.  To probe membranes with target bacterial cells, wash your membranes with PBST to remove salt... on the membrane to bind their target cells, pulling them out of solution. Wash your membranes well with... 20 Petri dish-sized circles of nitrocellulose membrane and press each gently down onto a plate of colonies...regrow the lifted colonies overnight), flip the membrane over and place it colony-side up onto a fresh ...plasmid selection. Remove air bubbles between the membrane and the agar using forceps. Several protein induction...

  18. Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)

    Type
    Blog Post
    ...multicomponent assembly, as fragments with overlapping sequence homology can be assembled in a unique order...multicomponent assembly have 5′ or 3′ sequence homology to each other, they may be assembled incorrectly.... one option is to perform a hierarchical assembly, assembling fragments in multiple steps to avoid using...Golden Gate assembly, may also be beneficial. SLIC is most often compared to Gibson assembly, a cloning...required for Gibson assembly. SLIC is a standardized method for multi-fragment DNA assembly, and its low cost... by Mary Gearing in December 2015 and was updated by Emily P. Bentley in September 2024.  References ... cheap, standardized, and rapid multi-part DNA assembly method. Read on to learn how to use it in your...

  19. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ...to use some assembly methods such as Golden Gate or Gibson assembly. Golden Gate Assembly of gRNAs Golden...Golden Gate/Gibson Assembly Multiplexing Plasmids: These plasmids allow you to assemble 2-4 gRNAs through...gRNAs. PTGs are assembled into a wt Cas9-containing vector using Golden Gate assembly, and up to 8 gRNAs...Multiplex Plasmid: Two gRNAs can be assembled using Gibson Assembly or SLIC cloning. gRNAs are expressed...Bullock lab, and gRNAs can be inserted using Gibson Assembly or SLIC cloning methods. A BsaI-based E. coli ... multiplexed gRNAs with Golden Gate and Gibson assembly methods If you want to scale up the number of ...Golden Gate Assembly uses Type IIS restriction enzymes, which cleave outside of their recognition sequence...

  20. Plasmids 101: Golden Gate Cloning

    Type
    Blog Post
    ...can be used to assemble multiple fragments - up to 10 fragments are commonly assembled in a single reaction... Gibson assembly require 20-40 bp of homology at the ends of DNA fragments to specify assembly order, ...sequence homology cannot be assembled using this method, but can be assembled with Golden Gate. The popular...acids, but Golden Gate assembly can be designed to be scarless. Golden Gate assembly is also less expensive...recognition sequence, they can be customized to direct assembly of DNA fragments. When designed correctly, the...sites with complementary overhangs that direct assembly of the final ligation product. As shown below,...higher number of potential clones. Golden Gate assembly has a few advantages over other cloning methods...
Showing: 61 - 80 of 629 results