...less efficient at taking up larger plasmids. If you need to transform large plasmids, it is a good idea...very few if any transformants when transforming large plasmids (>10 kb) or BACs, what can I do? Chemically...

...pipette is an essential tool for nearly anyone in molecular biology. You’ll use a pipette in many scenarios... liquid. Pro-Tip If pipetting from a relatively large container holding a small volume of liquid, tilt... of and gently in contact with the container (capillary action between the container and the liquid will...

...Antibody Plasmid Collection Antibodies Blog Posts Molecular Biology Reference General Transfection Protocol...and reagents from other vendors should produce similar outcomes when using these protocols. However, please...replication use a HEK293 line stably expressing the SV40 large T antigen. For antibody-expressing plasmids containing...

... an intravenous isomolar contrast agent. In research, iodixanol is used as an isomolar density gradient...60% iodixanol step Carefully add up to 5 mL of clarified supernatant on top of the gradient. Use 1X PBS...

...Kit-Free DNA Purification Agarose Gel Purification Molecular Biology Reference Introduction This protocol describes... (Sarkosyl) 0.1 M 2-mercaptoethanol TRIzol® or similar product such as TRI Reagent®, RNAzol®, QIAzol® ...Homogenize or lyse tissues or cells in TRIzol® or a similar product. For tissues: use 1 mL of TRIzol® per 100...

... Pooled Libraries All CRISPR Pooled Libraries Molecular Biology Reference How do I process my Addgene ...libraries up to 200,000 individual plasmids. For larger libraries scale the number of cells and DNA accordingly... LB Agar + Antibiotic 245 mm bioassay plates (Molecular Devices, X602) Pro-Tip Pour these plates at least...

...Information During several different stages of molecular cloning, it is important to get a quick and accurate... a liquid to determine the concentration of a particular substance in that liquid. Molecules absorb different...

...also use a nucleotide-based molecular insert (inosine) instead of a regular nucleotide for broader pairing...

... plate, do NOT dig into the agar. Another very popular technique is to draw in discontinuous lines. Start...30 °C instead of 37 °C. This is often true for large unstable plasmids, which sometimes recombine at ...

... pH7.5-8.0, 50mM NaCL, 1mM EDTA) and mixed in equimolar concentrations. We recommend mixing 2μg each ...μl). Mix the annealed oligos with cut vector in molar ratios (vector:insert) between 4:3 and 1:6 in a ...

... like... Cloning Methods Empty Backbone Guide Molecular Biology Protocols Overview Ligation Independent...Transform Mix your treated vector and insert at a molar ratio of 1:2 or 1:3, using between 20 and 50 ng ...

...success in numerous fields such as microbiology, molecular biology or cell biology. Learn some of the key...samples. This is a commonly used technique for molecular cloning, such as PCR- or restriction enzyme-based...

... a Liquid Bacterial Culture Introduction Many molecular biology techniques require highly purified and... of bacteria to grow (low copy plasmids require larger cultures). Centrifuge the culture to pellet the...

...will need to use a phosphatase to prevent re-circularization of the recipient plasmid. You should treat ...Additionally, if your final product is going to be very large (>10kb) you might want to use electro-competent ...

...from academic viral vector cores. Our titers were similar to those reported by these institutions. This protocol...Concentration of Addgene plasmid #59462 : 1.07 μg/μL Molecular Weight: 6208 bp x 650 dalton/bp or (g/mole)/bp...

...and reagents from other vendors should produce similar outcomes when using these protocols. However, please...plate washer (optional) Scale Microsoft Excel or similar software Reagents Purified antigen Purified recombinant...

...empirically, and that the cell population be monitored regularly for transgene expression. This protocol has been...over-confluent, trypsinize the cells and expand them to larger culture dishes, being sure not to cross contaminate...

...37°C, 5% CO 2 overnight. Although cells should regularly be passaged in DMEM + 10% FBS with penicillin/...limited to the implied warranties of fitness for a particular purpose, non-infringement, typicality, safety...

...places. Pro-Tip If you’re weighing out an amount larger than the capacity of your balance or weighing boat...

...cells for mycoplasma (Link opens in a new window) regularly Do not over- or under-grow your cells. Thaw a ...