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We narrowed to 967 results for: Cre

Showing: 161 - 180 of 967 results

  1. Plasmid Cloning by PCR (with Protocols)

    Type
    Protocol
    ...temperatures, but you may need to increase your primer length and increase the Tm if you are trying to clone...Bacterial Transformation Summary PCR based cloning is incredibly versatile and allows for nearly any piece of ... 2 ligations for each plasmid you are trying to create, with varying ratios of recipient plasmid to insert...

  2. Protocol - How to Streak a Plate

    Type
    Protocol
    ...of the plate, as shown in the diagram above, to create streak #1. Pro-Tips Hold your tooth pick at an ...bacteria over a second section of the plate, to create streak #2. Using a third sterile pipette tip, toothpick...bacteria over the last section of the plate, to create streak #3. Incubate plate with newly plated bacteria...

  3. Weighing Reagents Protocol

    Type
    Protocol
    ...using weighing paper, fold it in half first to create a crease and then place it unfolded onto the balance... sure you’re weighing all reagents precisely to create the solution you expect. Understanding how to obtain...

  4. Colony Formation Titering Assay

    Type
    Protocol
    ...likelihood of multiple integration events decreases as the dilution increases. Calculate the transduction units... remove cells and debris. Lentiviral titer can decrease during cycles of freeze-thaw. If you are freezing...

  6. Protocol - Bacterial Transformation

    Type
    Protocol
    ...Scientists have made many genetic modifications to create bacterial strains that can be more easily transformed... specific treatments have been discovered that increase the transformation efficiency and make bacteria...

  7. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ...it contains a high percentage of glycerol that increases the density of your DNA sample causing it settle...better resolution of bands? A few simple ways to increase the resolution (crispness) of your DNA bands include...

  9. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ... as opposed to transiently-transfected cells, increases reproducibility, as it eliminates the variation... vectors and genome-wide libraries for CRISPR screening. Sanjana NE, Shalem O, Zhang F. Nature Methods...

  10. CRISPR Guide

    Type
    Collection
    ...used for large scale edits. These proteins, like Cre recombinase or phage derived serine integrases, insert... GAT; increase nuclease fidelity SpCas9-NG - NG; increase in vitro activity SpG - NGN; increase nuclease...others work to increase Cas9’s proofreading capabilities. No matter the method, increased fidelity enzymes...backbone HypaCas9 - increase Cas9 proofreading and discrimination evoCas9 - decrease off-target effects...domains to expand PAM recognition, increase specificity, and decrease off-target effects Sniper-Cas9 - ...compatible with truncated gRNAs to increase specificity SuperFi-Cas9 - increases fidelity with reduced nuclease...integration of donor DNA. Genome-Wide Screens Using CRISPR In a genetic screen, a population of cells with mutations...

  11. Pouring LB Agar Plates

    Type
    Protocol
    ...maximum of ~100 colonies Pro-Tip If you will be screening a large number of colonies, we recommend using...your antibiotic to the molten gel mix, you should create a 1000x stock solution. For example: If you’ll ...

  12. Antibody Validation Using the Indirect ELISA Method

    Type
    Protocol
    ...an antibody. This protocol will describe how to create a standard curve using known concentrations of ...be empirically determined. The dilution series created in section 1, step 2 is a suggested starting point...

  13. AAV Production in HEK293 Cells

    Type
    Protocol
    ...harvesting. The addition of sorbitol has been shown to increase viral titers by up to 1.8-fold ( Rego et al., ...sodium bicarbonate, sorbitol has been found to increase viral titers by up to 1.8-fold. See the recipe...

  14. Plan Your Experiment

    Type
    Collection
    ... mutant allele of a gene (point mutant)? Increase or decrease expression of a target gene? Once you have...High-fidelity Cas enzymes increase specificity Dual-nickase approach increases specificity but is less ...efficiency. In fact, cleavage efficiency may increase or decrease depending upon the specific nucleotides ... the general framework of a CRISPR experiment. Created with BioRender.com. Select Your Desired Genetic...mammalian cell lines Interfere Activate (CRISPRa) Increase gene expression without permanently modifying ...likely won’t be able to completely avoid them. To increase specificity, consider using an engineered Cas ...target in your samples to ensure there are no discrepancies with the reference sequence you used for gRNA...

  15. Immunology Research Plasmids and Resources

    Type
    Collection
    ...Upon activation, NK cells secrete perforin and granzyme B, two enzymes that create pores within an infected...LSIGN, MGC129964, MGC47866 CREB1 cAMP responsive element binding protein 1 CREB, MGC9284 CTSB cathepsin ...chromogranin A (parathyroid secretory protein 1) CGA CHGB chromogranin B (secretogranin 1) SCG1 CKLF chemokine-like... SDS, SWDS SCG2 secretogranin II (chromogranin C) CHGC, SN, SgII SCGB3A1 secretoglobin, family 3A, member...HIN1, LU105, MGC87867, PnSP-2, UGRP2 SCT secretin - SCTR secretin receptor SR SCYE1 small inducible cytokine... B cells mediate humoral immunity through the secretion of antibodies, and also function as antigen presenting...

  16. CRISPR Plasmids - Base Edit

    Type
    Collection
    ...Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs...double stranded breaks. Cytosine base editors are created by fusing Cas9 nickase or catalytically inactive...converted to thymidine through base excision repair, creating a C to T change (or a G to A on the opposite strand..., which is treated like guanosine by the cell, creating an A to G (or T to C) change. Adenine DNA deaminases...deaminases do not exist in nature, but have been created by directed evolution of the Escherichia coli TadA...evolved TadA domain is fused to a Cas9 protein to create the adenine base editor. Both types of base editors...adjust the editing window, or adding fusions that increase product purity such as the DNA glycosylase inhibitor...

  17. CRISPR Pooled gRNA Libraries

    Type
    Collection
    ...In a CRISPR screening experiment, target cells are treated with the pooled library to create a population...mutant cells that are then screened for a phenotype of interest. Screening experiments using a pooled ...CRISPR pooled libraries for large-scale functional screens. Available for knockout, activation, inhibition...Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs...Blog Posts CRISPR is a useful tool for genetic screening experiments, due to the relative ease of designing...libraries and keep the following in mind: CRISPR screening experiments require electroporation to amplify...NGS) to confirm library representation. CRISPR screening experiments require large quantities of cells ...

  18. CRISPR History and Development for Genome Engineering

    Type
    Collection
    ...with work directed towards increasing on-target editing rates and decreasing off-target editing rates. ...others. Homology-directed repair (HDR) allows us to create a variety of modifications in endogenous loci with... locus makes it especially amenable to genetic screens. Pooled gRNA libraries can be used to identify ...sequencing, CRISPR is a robust system for genome-wide screening. CRISPR Technological Advances CRISPR technology... : Base editors directly convert bases without creating a double-strand break. This method displays higher...method targets RNA rather than DNA, it does not create permanent genome edits or double-strand breaks....epigenetic modifiers can modulate transcriptions and create heritable epigenetic marks. Visualize : Fluorescently...

  19. Plasmids for Stem Cell Research

    Type
    Collection
    ...reprogramming of adult pancreatic exocrine cells to beta-cells. Nature. 2008 Aug 27. Melton Pancreatic Acinar Cells...human cells in 2006 and 2007, respectively. To create iPSCs, fully-differentiated adult somatic cells...Plasmids in the table below. Once iPSCs have been created they can be directly differentiated into specific..., and transdifferentiation utilize plasmids to increase expression of key factors. The tables below will...plasmids available from Addgene that can be used to create iPSCs or another cell type. You can also search...iNDI) Collection for plasmids that can be used to create cell lines with endogenously-tagged gene variants...Plasmids Browse the table below for plasmids used to create induced pluripotent stem cells (iPSCs). Sort the...

  20. CRISPR Plasmids - Mammalian Expression

    Type
    Collection
    ... in the DNA. Indels often lead to frameshifts, creating loss of function alleles. To introduce specific...Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs...adenine to inosine, which is replaced by guanosine to create A->G (or T->C on the opposite strand) mutations...hypermutation in a 100 bp window. This technique creates diverse populations of mutants for directed evolution...breaks in DNA instead of the double-strand breaks created by wild type Cas enzymes. To use a nickase mutant...your DNA in close proximity. These double nicks create a double-strand break (DSB) that is repaired using...fused to a transcriptional activator peptide can increase transcription of a specific gene. Design your ...
Showing: 161 - 180 of 967 results