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...transforming large plasmids (>10 kb) or BACs, what can I do? Chemically competent cells are fast and easy to ...cell/DNA mixture to induce membrane permeability. To do this you will need to have access to an electroporator...

...μm) tissue sections and 3D cultures These samples do not necessarily require optical sectioning, but open...

...Updated Content last reviewed on 25 September 2025. Do you have suggestions for other plasmids that should...

...box and push the end of the pipette onto the tip. Do not touch the tips with your fingers to avoid contaminating...plunger, gently lower the pipette tip into the liquid. Do not submerge the pipette itself into the liquid and...

...regularly Do not over or under-grow your cells. Thaw a new vial of cells after 20–30 passages. Do not add...

...thicker as the proteins and DNA are denatured. Pro-Tip Do not vortex at this stage or the genomic DNA will ...4. Note: Phenol-chloroform is a hazardous waste - DO NOT pour down sink. Top Protocol: Ethanol Precipitation...

...Media 10 mL 10% Poloxamer 188 40 mL 200 mM Glutagro Do not add selective reagents Store at 4 °C until use...incubator on a shaking platform set to 120 rpm. Pro-Tip Do not use cells that are over 30 passages. Section ...

...adaptive immune functions that other CRISPR types do, due to a lack of canonical CRISPR features (Pinilla-Redondo...

...complementary to template strand). However, primers do not need to correspond to the template strand completely...

...scrape some of the frozen bacteria off of the top. Do not let the glycerol stock unthaw! Streak the bacteria...

... a new or experienced reprogrammer? Which methods do you have experience using? Let us know about your...

... significant medical interest, however efforts to do so have been hindered by a lack of tools, compounds...target soluble kinase domains, but these domains often do not express well in E. coli without their partner...

...surface. Tare the weighing boat or paper. You need to do this because you don’t want to include the weight...

...entire dilution plate with Microseal adhesive seal - do not press the film, just gently cover so nothing ...collecting drops along the side of the tube. Cap gently - do not push the cap in all the way, just ensure the ...

...the surface). We use the micropipette tip itself to do the spreading; the tip is gently bent to create an...

...of the tube. This way, as items are floating, you do not need to necessarily maneuver the bottles or tubes...

... 2025. Catalog items are updated more frequently. Do you have suggestions for other plasmids that should...

...Agudelo et al. prevent ouabain binding to ATP1A1 but do not compromise the ion pump’s function or cause cellular...

...the vector with EcoRI and SalI (see diagram ). To do this, we add 5' - AATTC and G - 3' to the top oligo...

...Protocol Storing and Handling Addgene Plasmids What to do after you receive your plasmids from Addgene Instructions...