...extracted the DNA and processed the mixed sample in our pipeline (see Figure 2, sample 2). When we blindly... potential contaminants. The analysis is a 2-step process using Geneious software: First, the individual...quality, we undertake a rigorous quality control process. In 2016, we launched our viral service to continue...bacteria snuck into the prep during the production process? These questions are especially important when ...CHRM4 gene. When contamination is suspected, we proceed to de novo assembly of the unmapped reads, employing...answer your questions about our quality control process! The images in this blog post were created using...

...neoblast cells and the gene Prohormone convertase 2 (pc2) labels the central nervous system. Evaluating... characterized in an unrelated worm (C. elegans) [2]. Delivering dsRNA matching a specific mRNA sequence...223). Link: http://abt.ucpress.edu/content/79/3/208 2. Fire, S. Xu, M.K. Montgomery, S.A. Kostas, S.E. Driver... its expression. More specifically, during the process of RNAi, an enzyme dubbed Dicer cuts or “dices”...Phagocata morgani and Phagocata gracillis, the process is a little more complicated. These strains are...RNA inhibits gene expression in planarians." Proceedings of the National Academy of Sciences 100.suppl...

... Plant Methods. BioMed Central, 2(1), p. 19. doi: 10.1186/1746-4811-2-19. PubMed PMID: 17087829. PubMed...PMID: 24032037. PubMed Central PMCID: PMC3759027. 2 Harrison, S. J. et al. (2006) ‘A rapid and robust ...from later. The Arabidopsis transformation process The process itself is straightforward. When your agrobacterial...growth on antibiotic-containing media) A similar process occurs in nature; Agrobacterium tDNA insertion ...in hand, so it is pretty important to get this process right the first time. I’ll lay out some guidelines...Agrobacterium strain. Additionally, the transformation process can get a bit messy, so you want to make sure you...begin working in the T2 generation. The selection process itself can affect the growth of resistant plants...

...CcdB.” Biotechniques. 1996 Aug;21(2):320-3. PubMed PMID: 8862819. 2. Bahassi, EM., et al. “F plasmid CcdB... desired insert should not grow.   Figure 2: Image from http://teachline.ls.huji.ac.il/72682/tutorials...still express ccdB and therefore would die. This procedure dramatically reduces the number of clones that...recombinant plasmid and therefore makes the cloning process much more efficient, as one does not have to thoroughly...

...indicated they wanted to see more negative data (Figure 2.) At the same time, when scientists who support open...is not only housed, but encouraged.    Fig. 2: Results from a survey conducted by Addgene in 2021...detailed methods and image descriptions, a minimum n = 2, and well labeled high- and low-magnification images...gene therapy trials. Human Gene Therapy Methods, 26(2), 71–76. https://doi.org/10.1089/hgtb.2015.040 Initiative...expression can take months (or longer), yet the process is often not detailed in the primary literature...

...vector:insert ratio is used in the DpnI digestion and 2 µL of digest product are used in the transformation...Biology 38 (2016): 145-154. PubMed PMID: 27391134. 2. Li, Chaokun, et al. "FastCloning: a highly simplified...in vivo. Detailed FastCloning procedure To start the FastCloning process, the insert and vector are first...oldest of these techniques and is a multi-step process consisting of digestion, purification, ligation...techniques with higher efficiency and simpler procedures have been developed. Some of these include, TA...specific sequences. Read on to learn how easy this process is and to get tips for applying it in your own ...Ligation + - Transformation + + The FastCloning process in brief In FastCloning, you PCR both the vector...

...NGG)  Editing window ~2 nt  85175  pBK-EE-BE3  SpCas9 (NGG)  Editing window ~2 nt  85176  pBK-YE2-...quantum yield of 0.54) and mScarlet-H with M164H (2-fold improvement in photostability compared to mScarlet...SpCas9 base editors with editing windows as small as 1-2 nucleotides! To reduce off-target effects associated...pBK-YE2-BE3  SpCas9 (NGG)  Editing window ~2 nt  85177  pBK-YEE-BE3  SpCas9 (NGG)  Lower on-target activity...activity than BE3; editing window ~1-2 nt  87438  pET42b-HF-BE3  HF-Cas9 (NGG)  Very low off-target activity...Ladders Are Awesome Tools: They are comprised of 2 plasmids, pPSU1 and pPSU2, that can be amplified affordably...libraries The Gross Lab at UCSF recently deposited 2 Bacillus subtilis libraries with Addgene – the B. ...

...various components of the immune system. Figure 2: In-vitro models for immunogenicity prediction of ...drug reaction. This system would somewhat act like a 2-step verification. All approaches have their own limitations...Future Enhancements. Frontiers in drug discovery, 2, 952326. https://doi.org/10.3389/fddsv.2022.952326Joubert...approach cannot replicate the complexity of antigen processing events that take place within an antigen-presenting...

...drastically with lower titers. As you can see in figure 2, the expression from the 5x1011 titer was essentially... roughly 6-fold increase in expression [3, Figure 2]. Note how expression increases exponentially with...international 61.1 (2002): S9-S15. PubMed PMID: 11841606. 2. Schultz BR, Chamberlain JS. Recombinant Adeno-associated...healthy as can be to prepare them for the arduous process of generating viral particles. Limit tube transfer...

...the light-inducible heterodimerizing cryptochrome 2 (CRY2) and calcium and integrin-binding protein 1 ...inactive, dead Cas9 protein (dCas9) fused to CIB1; and 2) the activator - the CRY2 photolyase homology region... fused to both the N- and C-termini of dCas9; and 2) CRY2FL-VP64, a fusion of full-length CRY2 and the...researchers saw an 11-fold increase in mRNA levels after 2 hr and a 400-fold increase after 30 hr. The system...authors fragmented Cas9 into N-terminal (residues 2-713, N713) and C-terminal (residues 714-1368, C714...nMagC714 instead of nMagHigh1-C714, generating paCas9-2. This change did not significantly alter the system...prior work, the Sato lab also showed that the paCas9-2 system could be spatially controlled and reversibly...

...non-specific proteins, and then elute YFP.  Option #2: Opto-Nanobodies (OptoNBs) purification You skip adding...blue lights. Which option do you choose?  Option #2 may sound like fiction, but light-controlled protein...experiments.  AsLOV2 Inertion Site mCherry #1 mCherry #2 GFP Loop 1 dark-induced dark-induced dark-induced...with a flip of a light switch would improve this process. The team used nickel-coated agarose beads coated...

...research experiences to undergraduate students (1, 2), but educators face significant challenges in designing... by western blotting in other experiments (Figure 2).   Complementation reveals conservation of function...Biology Education: A Call to Action, Washington, DC. 2. President's Council of Advisors on Science and Technology... controlled by the GAL1 promoter. The cloning procedure adds both V5 and His6 epitope tags to the C-termini...successfully analyze Met protein complementation. In the procedure diagrammed in Figure 3, students first transform...

...typically, normal (bell curve) or skewed (see Fig. 2). Assumptions – characteristics of a data set; ...levels of variance among data.     Fig. 2: Examples of data distribution profiles.   Parametric... positive is much higher than if you only look at 2 or 3 pairwise comparisons. Similar to family-wise ...statistical facets of reproducibility. Adv Physiol Educ. 40(2): 248-252 (2016). 0.1152/advan.00042.2016 Finch, W...Simulation Study. Front. Appl. Math. Stat. Volume 2 (2016). doi.org/10.3389/fams.2016.00002  ...Bonferroni Correction and the Benjamini-Hochberg procedure are designed to reduce false positive rates and...

...the result in Figure 1 could look like this: Figure 2: Expression of Plasmid B (as internal control) This...down expression of Gene X but, as shown in Figure 2, this was likely due to the original transfection ...PMID: 20074371. PubMed Central PMCID: PMC2817684. 2. Replicates and repeats—what is the difference and...transfection, transfection reagent, or transfection process) may have on the target cells. One transfection... the transfection reagents or the transfection process itself has any cytotoxic effects on the target ... optimization of the transfection reagents and process. It is important to note that optimal experimental...

...pH visible at the sample interface upon thaw (Fig. 2). Before thawing, all the samples were at the control...showed that AAV infectivity was not altered by a short 2-hour exposure to pH as low as 2.5 (Potter et al., ...Methods. 2013 Apr;10(4):278-9. PubMed PMID: 23538862. 2. Potter M, Lins B, Mietzsch M, Heilbronn R, Van Vliet...always looking for ways to improve our shipment processes. After reading a publication describing how short...

...digestion of the destination vector with SapI yields 2 sites- the first site accepts the sgRNA target sequence...marker, and connector to be released (Fig. 1, Step 2-3). Since donor plasmids of the same type will produce... a gRNA vector that targets the gene of interest, 2) a plasmid to specify the reading frame of the insertion...scientists have been able to study many cellular processes, including mitosis, mitochondrial fission/fusion...insertions can only be detected by labor-intensive processes like PCR or by evaluating visual phenotypes. Improving...

...3 plates at different densities for selection). A 2% agarose solution is then melted (and thereby sterilized...96-well assay plate. We generally freeze at least 2 ampules each of 4 clones with different levels of ...of agarose, which is then overlaid with the firmer 2% agarose gel solution discussed above for picking.... Protocol Pages Agarose cloning method procedure The procedure (illustrated in the YouTube video below...

... will repeatedly nose-poke at the active hole [1, 2].    Place preference/place aversion also models reinforcement...spend most of its times in the opposite chamber) [2]. Laser activation is usually controlled automatically...two different ways 1) your hypothesis is wrong or 2) there is some problem with the technical implementation...PMID: 21716290. Pubmed Central PMCID: PMC3775282. 2. McDevitt, Ross A., et al. "Serotonergic versus nonserotonergic...neuronal activation and is a sufficient readout to proceed with your behavioral experiments. Comparison of...

... is also gaining a lot of new traction lately (1, 2), depends on knowledge of the structure(s) a given...per plate) across 20 fresh selective plates. Step 2: Express random phage DNA fragments in E. coli Now...microbiology 23.4 (2015): 185-191. PubMed PMID: 25708933. 2. Cooper, Callum J., Mohammadali Khan Mirzaei, and ... biology 34 (2015): 69-77. PubMed PMID: 26275959. 2. Weigele, Peter, and Elisabeth A. Raleigh. "Biosynthesis...useful for the biotech industry, which is in the process of capitalizing on phage RBPs as diagnostic tools...DNA fragments with live host cells Now you can proceed with incubating this membrane with your cells of...

...   Figure 2: High-level overview of an indirect immunofluorescence procedure with key considerations... several steps outlined below and in Figure 2. For a deeper dive into the steps and techniques, see Im...recognize the species of your primary antibody and 2) is conjugated to a fluorophore that suits your experiment...primary antibodies is a time consuming and expensive process. Consider the vast number of possible primary antibodies...Performing an immunofluorescence experiment The process to perform IF can be broadly divided into the several...specific 3-dimensional epitopes. Different sample processing steps can alter epitopes in different ways, so...secondary antibody you use, so as long as the sample processing steps used in a non-fluorescent IHC/ICC assay...