...partitioning system in the bacteria, but generally it consists of a centromere-like region on the plasmid, a ...2015) Imaging centromere-based incompatibilities: Insights into the mechanism of incompatibility mediated...

...CRISPR/Cas Toolkit for plants includes 95 plasmids consisting of CRISPR/Cas nucleases, base editors, gRNA backbones...activation-based norepinephrine (GRABNE) sensors allow for sensitive and high spatiotemporal measurements of norepinephrine...

...technical roles. Therefore, institutions are responsible for ensuring that either all affected scientists...and Brandeis universities have stated that they consider postdocs paid on stipends to be FLSA exempt, and...

...bleach. Safety Warnings Lentivirus is generally considered biosafety level 2+. Please ensure that you are... media and 11.1 µg/mL Polybrene. Mix the cell suspension well before seeding. Mix each well with a 1 mL...45° angle. Count to 20 while slowly and gently dispensing the droplets down the side of the tube. Place... 95 10 2 1 Denaturation 94 0.5 2 40 Annealing/Extension 60 1 2 40 Enzyme Deactivation 98 10 2 1 Hold 4...

...have been described--CPEC (circular polymerase extension cloning) and SLiCE (Seamless Ligation Cloning ...multiple fragments, or synthetic biology, you should consider giving it a try! References: 1. Gibson DG, Young...

...this interaction will give you an idea of how responsive and supportive the ELN team is. Once you’ve made...information. If an employee leaves the institution, the transition is smooth as all data is organized within an ...

...great model organism because they are easy and inexpensive to grow and maintain in the lab, have a short... worm, Caenorhabditis elegans, has been used extensively as a model organism since the 1970s when Sydney...

...transgene expression from split vectors has been inconsistent. A greater understanding of basic AAV biology...vector strategies and lead to greater and more consistent transgene expression. References: Allocca M...

... Summers, 1998). Multimeric plasmids are also considered to be less stable and more prone to being lost...the supercoiled plasmid monomer, and at <6 kb, consistent with the size of a supercoiled plasmid dimer....

...into the mouse zygote (Qin et al., 2015).  When inside the zygotes, the gRNA will seek out its target ... Ong ST, Bergstrom DE (2018) Viable Mice with Extensive Gene Humanization (25-kbp) Created Using Embryonic...

... that have negative impacts on fitness. One consideration when making a transgenic cell line with the ...the piggyBac™ system, these challenges can be considerably reduced. The piggyBac™ transposon system can...

...demonstrated that selection for repair of a temperature-sensitive pha-1 mutation significantly enriches for knock-in...from a single injected animal.  PAM sequence considerations For changes 5’ to the PAM, sequences corresponding...

...training. Similarly, there are several factors to consider when selecting a postdoc mentor and lab. These... having sufficient data to write a manuscript. Consider questions like: What figures are missing? For ...

...human proteins, this would be time-consuming, expensive, and not to mention frustrating. One elegant solution...transfection, add csRNP and ssODN to the cell suspension, transfect and let cells recover for 2-5 days.Note...

... in the lab by heating to 56 °C for 30 min. Considerations Before You Start The health of the target cell...pipetting or inverting. Aliquot 1.35 mL of the cell suspension into each well of the 6-well dish directly on... 48–72 h. Pro-Tip Different cells may be more sensitive to lentivirus or to polybrene, in which case this...

...contacting [email protected]. These deposits are considered large deposits and are exceptions to our standard...and Christina Mork in February 2024.  Additional Inside Addgene Content on the Blog Quickest Way to Deposit...

...successfully [10-12]. The next stage of reprogramming consists of redifferentiation of iPSCs into the cell type...pluripotent stem cells (iPSCs) by temperature-sensitive Sendai virus vectors. Proc Natl Acad Sci U S A...

... In research, iodixanol is used as an isomolar density gradient medium suitable for virus purification...to mammalian cells. Therefore, and unlike other density gradient media, it is reportedly not necessary ...steps are used to remove contaminants with lower densities, including empty capsids. The 60% step serves ...

..., be aware that some gloves may contain latex. Consider nitrile gloves as an alternative. For extra precautions...anyone else associated with Addgene, can be held responsible for your use of the information contained in ...

...satellite colonies will grow). It is, however, more expensive. With a 6 cm diameter petri plate containing solidified...Plate 50 µL of transformed E. coli /rescue media suspension onto the agar and gently spread over the surface...