...best tips beyond the basics to get you started. 1) Be visible and open to connections This is a site... Also, many people on LinkedIn don’t appreciate random requests without a personal message and they may...trolls. People don’t like to connect with complete strangers as it borders on my first rule of networking--...a useful way to build or use a network. Total strangers can’t really help or refer you. Do not ask people... lot about requesting connections to complete strangers for informational interviews. Many have had success...considered intrusive and it’s still connecting with a stranger so is highly discouraged. You should be using ...

...manipulation, there are two main decisions to make: 1. Which modification? and 2. Which viral vector should... linked to a certain behavior. It is a very old branch of science, dating back several hundred years, ...genes. A popular method uses Cre recombinase on transgenic flox animals to introduce modifications at the...Timing: High expression levels of the viral vector trangenes typically occur two weeks after injection. Therefore... after several weeks which may also depend on transgene or promoter choice. Behaviors you can study using...Stoica L, Ahmed SS, Gao G, Sena-Esteves M. Gene transfer to the CNS using recombinant adeno-associated ...

...of interest and the metal ions.      Fig. 1: A sample containing a polyhistidine tagged protein...and several contaminating proteins (blue, green, orange) flows through the Ni2+-containing resin. The protein...polyhistidine containing contaminating protein (orange). A low imidazole wash buffer disrupts the weak...polyhistidine-metal ion interaction is not disrupted by denaturants (Hochuli, 1988). In some cases, proteins can ...tag and then refolded by gradual removal of the denaturant. Not all proteins will refold once denatured,...

...incorporated at the termini of the crRNA molecule (usually 1-3 of the first 3 bps are modified) for optimal stability... 3’ end. This repeat region associates with the trans-activating crRNA (tracrRNA) which interacts with... important cellular RNA molecules with post translational modifications. One of the most common of these...targeting experiments allows you to visualize transfection efficiency and even track localization in cellular...

...antibody for a…less sensitive moment.   Fig 1: Blugene fading into darkness because they accidentally...spilling over" into another channel's detection range. Though you can compensate for this when reading...

...advantage this cycle, consider the advice below:  1. Eligibility check Scrutinize every opportunity prior...increase your competitive edge, they alone, are no guarantee of success. Don’t luck into an internship, prepare...

...sites that were previously unavailable. Figure 1: The PAM sequence that the Cas nuclease recognizes...across a number of different sites. Using a combined random and directed mutagenesis approach, the Anna Cereseto...

...the output of the analysis performed in experiment 1. Use camelCase: In camelCase, each subsequent word...like a wet lab experiment I use the text editor TextWrangler to record the following, which are later printed...

...with each protein of interest.    Figure 1: The direct approach (left) versus the indirect approach...in light green; and proteins are represented in orange. Created with BioRender.com.    How do secondary...

... an entirely new affinity reagent.    Fig. 1: A comparison of an antibody (left) to several alternative...bonds, monobodies can be easily expressed in transfected eukaryotic cells, allowing for them to be used...an excellent option for many applications, and frankly, if ain't broke, don't fix it!  That being said...

.... See figure 1 for an example of how changing transfection ratios can affect transfection efficiency. ...containing structural genes and a transfer plasmid carrying the transfer gene. By dividing the viral elements... structural elements, envelope, and transfer gene are transfected into a packaging cell line, usually .... For some transgenes, transfection optimization simply is not enough to get a sufficient titer; this ...may not translate to the target cell line. To overcome this issue, we recommend using a range of doses...systems such that it no longer requires the viral transactivator, Tat. For more information on the different...requirement for additional DNA can impact the transfection efficiency of the packaging cell line and thereby...

...If you already have an end product in mind, steps 1 and 2 will generally be straightforward. Likewise ...reagents can be delivered via transfection, nucleofection, viral transduction or injection as either protein...cell line that can be transfected at high efficiency. Lentiviral transduction Cas9 and gRNA can be present... present in a single lentiviral transfer vector or separate transfer vectors. May contain reporter gene...inserted into an AAV transfer vector and used to generate AAV particles. Transient or stable expression...Chemical transfection   Lipid mediated Cationic polymers Calcium phosphate Cells Transformed cell lines... lines Transformed cell lines Transformed cell lines Cargo Nucleic acids or protein Nucleic acids Nucleic...

...patients lives. References 1. Ong-Abdullah, Meilina, et al. "Loss of Karma transposon methylation underlies...paternal grandmothers early food supply influenced cardiovascular mortality of the female grandchildren." BMC...histone modifications, which can influence the transcription of genes by RNA polymerase II, or by the addition...methylcytosine in CpG islands makes  genes less transcriptionally active. The role of epigenetics in studying...to play a large role in a variety of diseases, ranging from cancer, to diabetes, to congenital disorders...isolated population in Sweden found that paternal grandfathers who had experienced famine before their adolescence...adolescence passed on a heritable trait to their grandsons that decreased their likelihood of death from ...

...basis of many molecular biology tools! Figure 1: Common functions of natural plasmids. Created with...lateral”) gene transfer. Horizontal gene transfer can occur naturally through transformation (cellular uptake...? Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection Resources on Addgene.org...environment), transduction (viral infection by bacteriophages), or conjugation (direct transfer from one cell...one popular method scientists use to promote transformation, but it isn’t just for lab specimens — there... evidence that lightning can also promote gene transfer in soil bacteria! (Demanèche et al., 2001) Beneficial...humans only noticed last century! Nevertheless, the transfer of plasmids carrying these genes between bacteria...

... inhibitory synapses in the brain.  Figure 1:Schematic diagram of different FingR constructs and... efficient double stranded templates, rather than the more efficient single-stranded oligodeoxynucleotide...generally very small, and can contain different neurotransmitters. Using genetically encoded fluorescent proteins... them to localize to the synapse and minimize extraneous fluorescence from other regions of the cell. ...synaptic maturation pattern of hippocampal adult-born granule cells over time that were not previously possible... al., 2020). Figure 3: BaF3 cells were transduced with H2B-FT virus and grown in varying concentrations...lentiviral CRISPR library that targets human glycosyltransferase genes and other proteins related to cellular...

... – a mechanical device that could be used to bend 1/8th inch steel wire into a backbone structure of a...this surface”….where the complex is docked on the membrane of the endoplasmic reticulum. And graduate students...reported cleavage site on the complementary DNA strand. And this is why we love working with researchers...”. Jacob computationally added the non-template strand to the Jinek structure so that the resulting physical...protein must have in order to scan along the double-stranded DNA, recognize a PAM sequence, allow the complementary...pairing between the guide RNA and the target DNA strand, and then activate two different endonuclease active...structure captures the displaced non-target DNA strand and shows the HNH nuclease domain swung around ...

...article, they investigated three Casɸ orthologs, Casɸ-1, Casɸ-2, and Casɸ-3. This system is notable for several...including excision or insertion, inversion, translocation or cassette exchange. By utilizing recombinases...delivery constraints and scientists seeking a wider range of targetable genomic sequences could benefit from...spatially and temporally target single cells. Now, Francois St-Pierre’s lab developed a new form of FACS-based...-based Dual-fluorescent DSB Repair) is a double-strand break repair assay for mammalian cells. It's based...deposited by Leopold Parts. This library delivers 2 randomly paired guides per construct and allows a reduced...

... little signal detected along the neurite (Figure 1.) Moreover, the group was able to demonstrate that...expression in E. coli. This operon is under the transcriptional control of a strong constitutive promoter (... required enzymes, including a glutathione S-transferase homolog from a Janthinobacterium sp. Moreover... protein kinase A (PKA) activity at the plasma membrane in living cells using a construct in which TagRFP-T...used in quenching, FRET (Förster resonance energy transfer), and FLIM (fluorescence lifetime imaging microscopy...

...technologies (Bradbury & Plückthun, 2015a).   Figure 1: (Top) Polyclonal antibodies are produced in animals...chain and light chain for the antibody and when translated into protein will assemble into a fully functional...synthesize the genes that encode those amino acids (Tran et al., 2016). Each antibody would then be tested...P, Frenzel A, Wong J, Koh XY, Eng H-Y, Lane D, Lefranc M-P, Clark M, Dübel S (2018) When monoclonal antibodies...:633–644 . https://doi.org/10.1083/jcb.201409074 Tran NH, Rahman MZ, He L, Xin L, Shan B, Li M (2016) ...

...your finished constructs at Addgene!). References 1. Li, Chaokun, et al. "FastCloning: a highly simplified...would at least be guaranteed a successful result? While there are never any guarantees, some cloning techniques...Resources” section at the end of the post! Speed: Your grant deadline is approaching or your advisor is demanding...your insert, DpnI digest away the templates, and transform the PCR products into E. coli. Ligases within ...