...I’ve always been fascinated by the human body and how it works. I used to stay up past my bedtime, poring...baby’s health but adamantly refused to get them vaccinated. A classmate told me that they never finish a...more likely to rate that study as untrustworthy, inaccurate, methodologically flawed, and biased. As a result...information gained from that research remains inaccessible, behind paywalls and in technical jargon that...

...many cases, you won’t see any signs of DNA in your final tube after purification. How do you know if you ...your DNA prep and whether there are any other contaminants. Nucleic acids (DNA and RNA) absorb maximally...method also gives an indicator of DNA or RNA contamination based on the presence of other bands or streaking...methods, this method doesn’t tell you about contaminating proteins or chaotropic salts. Capillary electrophoresis...

...recent years, there are many fluorescent protein combinations that can be imaged together, but this also means...microscope(s) you intend to use. An accurate determination of whether two fluorescent proteins can be separated...Cy5 (e.g. Semrock's) and the similar 4-laser combination on a confocal (405 / 488 / 561 / 640 nm). In ...try several proteins in your system of interest. Finally, in this set of proteins the green and red proteins...

...backbone with a T7 promoter driving multiple combinations of WT, codon optimized, untagged, cleavage sites... sites (thrombin and TEV), and C-terminal tags (StrepII, 3xFLAG, 6xHIS, MBP, Myc). These constructs include...found in the bacterial expression plasmids. Each combination is present in two sets of backbones which drive...expression system. Mammalian expression plasmids Finally a set of wild type and codon optimized, full length...

..., but it’s also found in tissues. IgG is the predominant isotype during a secondary immune response, which...switching is caused by a specialized type of DNA recombination. Heavy chain constant genes are clustered together... Figure 2: Isotype switching requires DNA recombination of the antibody gene locus. B cells start by...heavy-chain constant domain gene cluster. DNA recombination can delete intervening heavy-chain constant ...

...continue to produce the specific antibody that the original B cell produced. Because of this, all of the antibodies...hybridomas producing the most specific antibodies. Finally, the selected hybridoma is expanded as a clonal...reproducible experimental results. When you are originally selecting a hybridoma, you can choose one that...from the hybridoma into a plasmid to create a recombinant antibody.     Figure 2: Changes to the...

...the page. Finally, as previously mentioned, enlist the help of your network to polish the final draft prior...don’t guess. Simply reach out to the program coordinator with your question(s). This precautionary step...notice within 24 hrs, send a note to the program coordinator.   6. Virtual interview prep Prepare for your...

...sides of this promoter-gene combo are two long terminal repeats (LTRs). These LTRs define the limit of...respectively, act as RNA polymerase II promoter and terminator regions and are slightly different in sequence...expression constructs. Sandwiched between two inverted terminal repeats (ITRs) is the promoter and cargo, in that... it to evade detection by the host. Of note, recombinant adenovirus is missing the endogenous E1 gene,...

...lot in common. In fact, selections are often inappropriately categorized as screens! Why the confusion? ...Alternative screening techniques While CRISPR and drugs dominate the screening scene, there are other options which.... Non-functional traits will be automatically eliminated, unlike in screens. Selections begin and end ...require there to be a culture condition which eliminates manipulations that do not affect the function...

...Inflammation and Immunity, as well as various combinations thereof (so called “Immuno-Oncology”) is exploding...tackle the challenge of curing cancer in cross-disciplinary projects and collaborations. Summarized by Professor...the boost it needs to help rid us of cancer in combination with chemotherapy. To dive deeper into this field...exciting development in immunotherapy is the combination of the CRISPR/Cas9 methodology with T cell receptor...

...antimicrobial.  Conjugates And HRP brings us handily to the final topic of this post, the conjugates, or signaling... a brief introduction to the wide, wild, and fascinating field of signaling molecules. Assay Type Westerns..., simple to use, not sensitive to light in its inactive state, and cheap. Unless there’s a specific reason... on Addgene.org Find plasmids encoding recombinant antibodies at Addgene Topics: Antibodies...

...well as an efficient mechanism of homologous recombination to facilitate simple gene replacement/mutation...directly into the host chromosome via homologous recombination. Yeast Replicating plasmids (YRp):  These vectors...some antibiotics. Auxotrophy is defined as the inability of an organism to synthesize a particular organic...selection element pairs must be utilized or a new combination needs to be created in advance of the experiment...

...convenient site for conjugation.  Finally, Fvs are the most amenable to recombinant expression and will have the...If so, you are not alone. Full-sized antibodies dominate the world of research affinity reagents, and for...not a “Fab” fragment, Fvs are often produced recombinantly and engineered to be joined by a linker peptide...

...repair mechanisms that can be exploited to either inactivate a gene or introduce a desired genetic alteration...expression. Its catalytic domains can be mutated to inactivate the protein’s nucleolytic capability, and this...remains intact while its nucleolytic activity is eliminated (Fig. 1) (1, 3). The gRNA can be further modified...tight control over Cas9 nuclease activity while eliminating the reliance on orthogonal Cas9 species. Within...

...secondary will bind to all the primary antibodies indiscriminately, giving them the same output signal. In these... Time is a precious commodity in the lab and eliminating the need for secondary antibodies would save ...use a primary antibody labelled with a probe, eliminating the need for a secondary antibody. Other immunohistochemistry... on Addgene.org Find plasmids encoding recombinant antibodies at Addgene  ...

...temperature, humidity, noise level, circadian effects (inactive versus active phase) Experimenter Interaction... of certain genes. A popular method uses Cre recombinase on transgenic flox animals to introduce modifications...- from basic types like locomotion and motor coordination to more complex situations such as learning ...Sena-Esteves M. Gene transfer to the CNS using recombinant adeno-associated virus. Curr Protoc Microbiol...

...research focuses on understanding the structural determinants of protein-DNA interactions and how these interactions...functional portion of the Nfix gene using Cre recombinase and to generate Nfix-negative neural stem cells...developmental delay, result in loss of function or dominant negative NFIX phenotypes. Developing a rapid ...

...post was originally written by Tyler Ford in March 2017 and updated by Angela Holmes and Christina Mork in..., spectinomycin, streptomycin, tetracycline, combinations of the above, or other. High or Low Copy... with highly repetitive sequences (prone to recombination in bacteria) such as lentiviral, retroviral,...

... proceeding to ultracentrifugation to avoid contaminating your transduction with the packaging cells. ...may be the result of plasmid integration via recombination during transfection, and does not necessarily...PMC84922. 5. Ugai, Hideyo, et al. "Stability of a recombinant adenoviral vector: optimization of conditions...., Michael J. Yezzi, and Barbara R. Young. "Recombination events during integration of transfected DNA...

... lab page, then onto the publication page, and finally on down to the individual plasmid page. The depositing...modifications listed can refer to changes made to the original backbone by the depositing lab, or by a previous...previously deposited with Addgene, you can list the original vector’s plasmid number here. Gene/insert: This...plasmid contains certain fluorescent proteins. Finally, for many plasmids you can find additional resources...