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

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We narrowed to 451 results for: abo.5

Showing: 421 - 440 of 451 results

  1. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    ...1087–1093 . https://doi.org/10.1038/s41592-019-0614-5 Sztal TE, Stainier DYR (2020) Transcriptional adaptation... “transitivity,” calls into question any claims about specificity of a given RNAi technique. While transitivity...conceptually similar to the microRNA precursor mentioned above and is a popular RNAi trigger in the mammalian research...substantial ways: variable silencing efficiency and labor-intensive processes to isolate RNAi plants. With...resource on the Addgene blog Browse our blog posts about plant biology Check out our CRISPR blog posts Resources...

  2. Antibodies 101: Antibody Engineering and Directed Evolution

    Type
    Blog Post
    ...autoimmunity and inflammation. Nature Reviews Immunology, 10(5), 301–316. https://doi.org/10.1038/nri2761 Chiu, M...structure-guided directed evolution. The nice thing about these methods is that they all follow the same general...performance. If you are interested in learning more about antibody engineering technologies, the Institute...

  4. Kit Free RNA Extraction

    Type
    Protocol
    ...cells. Allow sample(s) to sit at room temperature for 5 minutes to allow for dissociation of the nucleoprotein...Ethanol and vortex for a few seconds. Centrifuge for 5 minutes at 10,000 x g at 4 °C and remove the supernatant...without disturbing the pellet. Air-dry the pellet for 5-10 minutes. Critical It is important to not let the... working with the volatile reagents in the list above. Procedure Option #1 - Solution D Protocol Before...

  5. Viral Vectors 101: Optogenetic Tools

    Type
    Blog Post
    ...), 1–20. https://doi.org/10.1038/s41467-021-24759-5 Zhang, K., & Cui, B. (2015). Optogenetic control of...which tools to use, and what do you need to know about them to be successful in your experiments? In this...

  6. AAV Production in HEK293 Cells

    Type
    Protocol
    ...resuspended cell pellets and sonicate 5 x 1 sec pulses with at least 5 minutes on ice between each pulse,...resuspend the pellets in a total of 5 mL of cell lysis buffer (recipe above). Pipet back and forth to resuspend.... T-175 flask, Corning 430825, 175 cm 2 Cellstack 5, Corning 3319, 3180 cm 2 Cellstack 2, Corning 3269...high glucose, add 55 mL of heat-inactivated FBS and 5 mL of glutaGRO 11 mL of 200 mM L-alanyl-L-glutamine... water + 100 mL of 1 M Tris HCl pH 8.5 + 60 mL of 5 M Sodium Chloride + 4 mL of 1 M Magnesium Chloride....8-fold. See the recipe for D1 + 0.1 M sorbitol above. Carefully pour off the media into a waste container...pellets and keep on ice. Process the cell pellet from above as follows: Resuspend and lyse the cells by adding...

  7. Cancer and the Immune System: Deciphering the Relationship

    Type
    Blog Post
    ...PMID: 25998715. PubMed Central PMCID: PMC4491443. 5. Scott, Andrew M., Jedd D. Wolchok, and Lloyd J. Old...potential beneficiaries. Collaborative therapy For the most efficient outcomes, collaborative therapies must be... of the tolerance-producing antigens enunciated above. Finally, signals released from the microenvironment...are in their clinical trial phase. In all of the above therapeutics, techniques like genome engineering...are beginning to see the fruits of many years of labor with up and coming immunotherapy treatments. As ...clinic, physicians and researchers are learning much about how treatment regimens can be improved and how the...ahead of the disease in patients and developing collaborative treatments will go a long way in combating this...

  8. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    ...PloS one 4.10 (2009): e7338. PubMed PMID: 19823671. 5. Griffin, B. Albert, Stephen R. Adams, and Roger Y...environment? If these are pressing issues in your laboratory, then FRET experiments are a viable biophysical...to consider these cases separately when talking about the tools that are available. Let’s consider the...that, if you make an FP fusion, you are inserting about 20-25 kDa of protein mass into your protein and ...together The FRET donors and acceptors described above can be paired together to measure a wide range of...of your protein that can lead to novel insights about its biology and its behavior. See the table below...

  9. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

    Type
    Blog Post
    ...PMID: 21596592. PubMed Central PMCID: PMC3164316. 5. Chieppa, Marcello, et al. "Dynamic imaging of dendritic...fluorescence microscope where the light source is mounted above (epi) the specimen and the excitation light passes...technique that allows imaging of living tissue up to about one millimeter in depth. Two low-energy photons ...response, but the study of these dynamics was still laborious as epifluorescence microscopy wasn’t able to give...skin. Two-photon intravital microscopy. The video above depicts the infiltration of neutrophils expressing...

  11. Gibson Assembly Protocol

    Type
    Protocol
    ...can be created via PCR with primers that contain a 5′ end that is identical to an adjacent segment and ...single-strand DNA 3’ overhangs by chewing back from the DNA 5’ end. Complementary DNA fragments can subsequently... to several hundred kilobases. Nature Methods , 6(5), 343–345. https://doi.org/10.1038/nmeth.1318 (Link... the right). When designing your plasmid, think about what DNA segments you will need to join to create...

  12. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    ... CRMAGE is an incredibly fast protocol, with only 5 hours incubation time needed for a single round of...CRISPR Featured Topic Page Learn more about Cpf1 Find out about CRISPR Multiplexing Resources on Addgene.org...coli and other bacterial species. Read on to learn about the tools available for bacteria and some of the...that can be used in this way. As described in the above CRISPRi section, the Marraffini Lab published a ...as a collection of organisms, their genes, and metabolic processes, occupying the gut. Many human diseases...

  13. Antibody Validation Using the Indirect ELISA Method

    Type
    Protocol
    ...the wells. Immediately before use, mix 5 mL TMB Solution and 5 mL Peroxide Solution from the TMB substrate...at room temperature or overnight at 4 °C . Section 5: TMB reaction Carefully remove the plate seal from...point. If your unknown sample’s absorbance falls above the range of the standard curve you will need to...

  14. Technical Design of a Western Blot

    Type
    Blog Post
    ...over a broad dynamic range that typically spans 2–5 orders of magnitude, which includes a wide dynamic...If you’ve ever run a western blot, or thought about running one, you’ll know there’s a lot of choices...

  15. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...DMEM complete + 10 µg/mL polybrene (µL) 0 0 500 1:5 300 200 1:10 150 350 1:50 30 470 1:100 15 485 1:500...well gets one dilution, so a 6-well plate will hold 5 dilutions plus one 'no virus' control well). Perform...cells in the untransduced well (0 µL lentivirus, above) are dying. Perform regular media changes and monitor...

  16. Protocol - How to Perform Sequence Analysis

    Type
    Protocol
    ...confirm. Addgene's plasmid information pages recommend 5’ and 3’ sequencing primers. These primers typically...Addgene lists the primers used to obtain each result above the posted sequence in the "View Sequence" link....sequence than expected and wish to contact Addgene about the accuracy of your plasmid, please email help@...

  17. Lentiviral Vector Guide

    Type
    Guide
    ...U5 in cis Unique 5'; in third-generation plasmids, this region is often removed in 5’ LTRs and replaced.... Second-generation plasmids include tat , as the 5’ LTR is used as a promoter on the transfer plasmid...Instead, the transfer plasmid contains a chimeric 5’ LTR fused to a heterologous promoter (often CMV or...self-inactivating (SIN). This deletion is transferred to the 5’ LTR after one round of reverse transcription, inhibiting...Transfer plasmid — contains transgene and LTRs (chimeric 5’ LTR) Packaging plasmid 1 — contains gag and pol Packaging...self-inactivation LTR Viral Promoter Wildtype Hybrid promoter; 5' LTR is partially deleted and fused to a heterologous...the lentiviral vectors from your plasmids (Figure 5). This process requires a production cell line, typically...

  18. Gamma-Retroviral Vector Guide

    Type
    Guide
    ...sequence. U5 in cis Unique 5'; in some newer plasmids, this region is removed in 5’ LTRs and replaced with... of the viral genome that is transferred into the 5' LTR after one round of reverse transcription. After...with a heterologous promoter (usually CMV or RSV). 5' LTR in cis Acts as an RNA pol II promoter; the transcript...transcript begins at the beginning of R, has a 5' cap structure, and proceeds through U5 and the rest of...of the provirus. Some newer plasmids use a hybrid 5' LTR with a constitutive promoter such as CMV or RSV...3' LTR in cis Terminates transcription started by 5' LTR by the addition of a polyA tract just after the...Read our gamma-retroviral guide to learn about gamma-retroviral components, retroviral production, and...

  19. CRISPR Library Amplification

    Type
    Protocol
    ... µL, 10 µL) Bacti Cell Spreaders (VWR, 60828-680) 5 mL and 10 mL Serological pipettes Ice slurry (Ice ... mL Vented Falcon Tubes should contain a total of 5 mL (3 mL SOC + 2 mL transformed Endura from two separate...require modifications dictated by the originating laboratory for optimal results. If you obtained the pooled...

  20. CRISPR Guide

    Type
    Guide
    ...bacterium (LbCpf1) 5' TTTV AsCpf1 RR variant 5' TYCV LbCpf1 RR variant 5' TYCV AsCpf1 RVR variant 5' TATV Campylobacter...sequence may not be characterized *In the table above, 3' and 5' indicate on which end of targeted sequence...will continue to anneal to the target DNA in a 3′ to 5′ direction. The location of any potential mismatches...inhibit target cleavage, while mismatches toward the 5′ end distal to the PAM often permit target cleavage...from naturally occurring cytidine deaminases. Figure 5: Overview of base editing More recently, researchers...transcription. The edited DNA strand displaces the unedited 5’ flap, and the resulting heteroduplex is resolved ... repair using the edited strand as a template PE4/5 - improved PE2/PE3 to inhibit mismatch repair to promote...
Showing: 421 - 440 of 451 results