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Showing: 1 - 50 of 137 results

  1. CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments

    Type
    Blog Post
    ...to adapt the popular guide design tool Crispor by Max Haeussler. With a few extra lines of code, we enabled...protospacer + PAM) in the target region after HDR. If the max CFD score is less than 0.03, it stops mutating. (...Zuckerberg Biohub Manuel Leonetti Ryan Leenay Jason Li Max Haeussler Luca Pinello Kendell Clement Andy May ...

  2. The Effect of COVID-19 on Liu Lab Plasmid Requests From Addgene Data

    Type
    Blog Post
    ...This post was contributed by Max W. Shen from MIT, Alvin Hsu Harvard University, and David R. Liu from... uncertainty. Many thanks to our guest bloggers Max W. Shen from MIT, Alvin Hsu Harvard University, and...from the Broad Institute and Harvard University. Max Shen is a Ph.D. candidate at MIT. His research uses...gradient descent in Pytorch, which gave us the maximum likelihood estimates of each parameter. We approximated...

  3. Advancing Biology with Zebrafish: Genetic Tools for Developmental Studies and More

    Type
    Blog Post
    ... researcher at the Friedrich Miescher Lab of the Max Planck Society. Zebrafish (Danio rerio) have been...Katherine Rogers from the Friedrich Miescher Lab of the Max Planck Society. Katherine W. Rogers is a postdoctoral...group at the FriedrichMiescher Laboratory of the Max Planck Society in Tübingen, Germany. She studies ...

  4. Bright Monomeric Fluorescent Proteins: mNeonGreen, mTFP1, and mWasabi

    Type
    Blog Post
    ...narrow excitation and emission peaks (max excitation at 493 nm and max emission at 509 nm), making it useful...GFPs and YFPs. Its excitation maxima is at 506 nm and its emission maxima is at 517 nm. Because mNeonGreen... to CFP484. Because the excitation and emission maxima were red-shifted (462 nm and 492 nm, respectively... mWasabi can be used with Sapphire (excitation maximum 399 nm) because it does not significantly excite...

  5. Visualizing Protein Turnover In Situ

    Type
    Blog Post
    ...is organized by international PhD students at the Max Planck Institute for Biophysical Chemistry, Göttingen...a former Caltech researcher, now Director at the Max Planck Institute for Brain Research in Frankfurt ...

  6. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    ...contributed by Sana Khan Khilji, a PhD student at the Max Planck Institute of Colloids and Interfaces. Here...Germany. She will soon be joining her PhD lab at the Max Planck Institute of Colloids and Interfaces. Sana...

  9. Transfection for Recombinant Antibodies

    Type
    Protocol
    ...store the solution at -20 °C. 1 mg/mL PEI-MAX Add 1 g of PEI-MAX powder to 900 mL deionized water in a 1... mg/mL PEI-MAX to the second tube of 6 mL BCD TFX (for a 1 mg/mL stock solution of PEI-MAX, 450 µg = 450...recombinant antibody plasmids using Polyethylenimine Max as a transfection reagent. After transfection and... Polyethylenimine hydrochloride, M.W. 40000 (PEI-MAX), Linear, Transfection Grade, VWR 75800-188 10% Pluronic...Before Starting Warm the DNA and working stock of PEI-MAX to room temperature before use, and warm the BalanCD...for 1 h in the 37 °C bead bath. Transfer the PEI-MAX and recombinant antibody plasmid DNA sample to the...ratios may range from 1:1 to 1:6. Add the diluted PEI-MAX to the diluted DNA. Cap the tube and vortex with ...

  14. An Addgene Summer Internship

    Type
    Blog Post
    ... safety reasons, and the insert size should be a max of 500 bp (preferably). I compiled a list of three...

  20. TALENs for Endogenous Zebrafish Genes

    Type
    Collection
    ...TGTTCCTGGTGGCGGTGCatgaattgggtcatgcaCTTGGTTTGGAGCACTCA max TAL3308 & TAL3309 TGAGCGACAACGATGAtatcgaggtcgacagtGATGTGAGTATAGTTA...

  22. Your Lentiviral Plasmid FAQs Answered

    Type
    Blog Post
    ... and we would be happy to answer them! Note: A. Max Juchheim, Marcy Patrick, and Meghan Rego contributed...the 3rd-generation packaging systems offer the maximal biosafety available for the technology. The minor...

  25. Lentivirus Production

    Type
    Protocol
    ...chloroquine. After the incubation, add the DNA:PEI-Max mix to the conical containing the media and chloroquine...

  26. CRISPR Library Amplification

    Type
    Protocol
    ...65 mm Petri dish (VWR, 11019-552) 4 MaxiPreps (Qiagen HiSpeed Max, Catalog #12663) Tips (1000 µL, 200 ... for the purpose of a Maxiprep. Purify plasmid DNA using the Qiagen HiSpeed Maxi Kit (one conical is its...representation is maintained. Maxipreps - Less is more: Do not overload the Maxipreps as yield can dramatically...spread out the harvested cells further than four Maxipreps worth. Count colonies on the most dilute Petri... its own Maxiprep). Critical Do not freeze pellets for later purification. Immediately purify them! Commercial...Commercial Maxipreps rely on incremental, ordered cell lysis. E. coli cells are subject to lysis by freeze-thaw...Glycerol or DMSO solutions. Quantify the individual Maxipreps by Nanodrop, Picogreen, or Qubit. Each different...

  27. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...Suppmann, the Recombinant Protein Production group at Max-Planck Institute of Biochemistry has developed a ...Multisite Gateway compatible destination vector for a maximum of 24 elements assembled into a final transgenesis...

  28. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...interaction strengths. Erich Wanker’s group at the Max-Delbrueck Center for Molecular Medicine has recently...independent gRNAs, each chosen strategically to maximize the likelihood of creating a functional knock-...

  29. Jaenisch Lab CRISPR Plasmids

    Type
    Collection
    ...dCas9VP64 on pmax expression vector. 48224 pAC92-pmax-dCas9VP96 : dCas9VP96 on pmax expression vector. 48225... 48225 pAC93-pmax-dCas9VP160 : dCas9VP160 on pmax expression vector. 48226 pAC94-pmax-dCas9VP160-2A-puro...promoters. Table 2. pmax dCas9 Activator Expression Plasmids ID Plasmid 48223 pAC91-pmax-dCas9VP64 : dCas9VP64...Plasmid 48222 pAC90-pmax-DEST : Gateway Destination vector derived from Clontech's pmax (CAGGS) expression...activators and sgRNAs can be separately transfected (pmax dCas9 activators [48223~48227] + sgRNA expression...transferred to Gateway destination vectors (e.g., pmax-DEST [48222] and (Link opens in a new window) Lifetechnologies...2A-puro : dCas9VP160-2A-puro (puro-selectable) on pmax expression vecor. Table 3. dCas9 Activators Gateway...

  30. Trimmer Lab NeuroMab Collection

    Type
    Collection
    ...Mouse IgG2a 114541 Anti-KCNU1/Slo3 maxi-K+ channel [N2/16R] KCNU1/Slo3 maxi-K+ channel Mouse Mouse IgG2a 114542...IgG2a 128634 Anti-Slo1/BKAlpha maxi-K+ channel [L6/23R] Slo1/BKAlpha maxi-K+ channel Mouse Mouse IgG2a ...IgG2a 177486 Anti-Slo1/BKAlpha maxi-K+ channel [L6/60R] Slo1/BKAlpha maxi-K+ channel Mouse Mouse IgG2a ...IgG2b 199406 Anti-Slo1/BKAlpha maxi-K+ channel [L6/60R-2b] Slo1/BKAlpha maxi-K+ channel Rat Mouse IgG2b 199407...IgG2a 206523 Anti-Slo1/BKAlpha maxi-K+ channel [L6/48R] Slo1/BKAlpha maxi-K+ channel Mouse Mouse IgG2a ...IgG2a 222180 Slo1/BKAlpha maxi-K+ channel [L6/60R-rat IgG2a] Slo1/BKAlpha maxi-K+ channel Mouse Chimera...Mouse 199421 Slo1/BKAlpha maxi-K+ channel scFv [L6/60] L6/60 scFv Slo1/BKAlpha maxi-K+ channel Rat Mouse 199422...

  31. CRISPR 101: Cytosine and Adenine Base Editors

    Type
    Blog Post
    ...indel frequency. Koblan et al., 2018 BE4max, ArcBe4max, ABEmax   Improved nuclear localization and expression...localization signals and codon usage BE4 to create BE4max and AncBE4max with a 4.2-6-fold improvement in editing...adenine base editing When the Liu lab created BE4max and AncBE4max mentioned above, they also made an adenine...., 2020 SpRY Cas9, available fused to ABEmax(7.10) or to BE4max   Evolved SpCas9 with NRN or NYN PAM site... more flexible sequence preferences, evoAPOBEC1-BE4max, which is significantly more effective at editing...localization and expression. This base editor was named ABEmax. In 2020, two papers were published describing ...programmable adenine and cytosine editor) by fusing miniABEmax-V82G and Target-AID to Cas9 (Grünewald et al....

  32. Prime Editing: Adding Precision and Flexibility to CRISPR Editing

    Type
    Blog Post
    ... continued to improve the maximum efficiency of prime editing. While maximum efficiency isn’t everything... PE2-5 approaches, sometimes referred to as PE2max, PE3max, etc. PE6: Specialized prime editing variants...end of PEmax to produce PE7. PE7 significantly improves editing efficiencies compared to PEmax with normal... originally introduced, rely on the PE2 enzyme. PEmax: Optimizing prime editor sequence to improve expression...be combined with any of the PE2-PE5 strategies. PEmax contains a reverse transcriptase with a sequence... although both enzymes still approach or exceed PEmax editing efficiencies for short, simple edits. PE6c... Innovation Purpose PE2 PE3 PE4 PE5 PEmax PE6 PE7 Pentamutant reverse transcriptase...

  33. Antibodies 101: Designing Your First Flow Panel

    Type
    Blog Post
    ...spectrum shows two maxima, one local maximum at just under 500 nm and a global maximum at around 560 nm,... global excitation maximum lies around 650 nm, APC-Cy7's global excitation maximum is about 750 nm. If...and emission spectra. Both spectra possess clear maxima where excitation and emission are the most efficient...fluorophores. Each fluorophore has a defined emission maximum. Note, you can also use the tool to display excitation...wavelength of 488 nm (Table 1), which is near a local maximum of PE's excitation spectrum (dotted line, Figure...excites at 561 nm, which is close to the global maximum of PE's excitation spectrum where the excitation...

  34. CRISPR History and Development for Genome Engineering

    Type
    Collection
    ... M, Rollins MF, Hidalgo-Reyes Y, Wiedenheft B, Maxwell KL, Davidson AR. 2015. Multiple mechanisms for ...136-9. PMID: 26416740 Bondy-Denomy J, Pawluk A, Maxwell KL, Davidson AR. 2013. Bacteriophage genes that...Hidalgo-Reyes Y, Lee J, Edraki A, Shah M, Sontheimer EJ, Maxwell KL, Davidson AR. 2016. Naturally occurring off-...

  35. CRISPR Guide

    Type
    Collection
    ... a structured motif to enhance pegRNA stability PEmax - optimized PE architecture; increased synergy between.... PMID: 25079318 Bondy-Denomy, J., Pawluk, A., Maxwell, K. L., & Davidson, A. R. (2013). Bacteriophage...Rollins, M. F., Hidalgo-Reyes, Y., Wiedenheft, B., Maxwell, K. L., & Davidson, A. R. (2015). Multiple mechanisms...Kranzusch, P. J., Sontheimer, E. J., Davidson, A. R., Maxwell, K. L., & Doudna, J. A. (2017). A Broad-Spectrum..., J., Edraki, A., Shah, M., Sontheimer, E. J., Maxwell, K. L., & Davidson, A. R. (2016). Naturally occurring...

  36. Fluorescent Protein Guide: Empty Backbones

    Type
    Collection
    ... 50% of its maximum value. Maturation is the time for fluorescence to obtain half-maximal value. Structure...Color) For each table: Excitation and Emission maximum are listed in nm. Brightness is the product of ...

  37. CRISPR Plasmids - Prime Edit

    Type
    Collection
    ...inhibit MMR PE5 - PE3 plus MLH1 mutant to inhibit MMR PEmax - optimized PE2 enzyme containing RT optimized for...RNA-binding exonuclease protection factor La to PEmax to enhance pegRNA stability epegRNA - additional...

  38. CRISPR References and Information

    Type
    Collection
    ...mulatta). Cas-Designer searches for targets that maximize knockout efficiency while having a a low probability...dual expression: pAC2 , pAC152 , pAC153 , pAC154 ; pmax dCas9 activator expression: pAC91 , pAC92 , pAC93...

  40. Quantifying DNA? Here are Five DNA Quantification Methods to Consider

    Type
    Blog Post
    ...molecules have a specific wavelength that they maximally absorb. The spectrophotometer measures these absorbances...contaminants. Nucleic acids (DNA and RNA) absorb maximally at 260 nm. Proteins on the other hand absorb best...nm and organic compounds and chaotropic salts maximally absorb at 230 nm. The A260/A280 ratio is used ...

  43. What's New in CRISPR - March 2020

    Type
    Blog Post
    ...deamination. They generated these editors using the BE4max architecture. The new editors reported in this ...average on-target DNA-editing levels relative to BE4max.  Read the paper in Nature Biotechnology Find ...

  45. What's New in CRISPR - November 2021

    Type
    Blog Post
    ...amino acid sequence of the prime editor, yielding PEmax, to work synergistically with epegRNAs and PE4 and...disease.  Read the paper in Cell Find PE4, PE5, and PEmax plasmids Precise genomic deletions In PRIME-del...

  48. Antibodies 101: Stripping and Reprobing Western Blots

    Type
    Blog Post
    ...through several rounds of stripping and reprobing, maximize your signal by starting with the least-abundant...Immediately after transfer, dry the membrane to maximize protein retention. Following this step, most PVDF...

  50. Hot Plasmids: Winter 2025

    Type
    Blog Post
    ...1: Left: recombined prime editing tools PE2max-NC and ePEmax3 optimized for dicots. NC: nucleocapsid RNA...
Showing: 1 - 50 of 137 results