...faster when using a dual guide system.   Fig. 2: Size screening for large deletion events.   ... Biol 17: https://doi.org/10.1186/s13059-016-1012-2 Find more CRISPR Resources at Addgene: Read our CRISPRGuide... as a proxy for your edit of interest. Using the same gRNA and donor, you can deliver a silent mutation...to compare the sequencing reads from your edited sample to an untreated population. Software such as CRISPResso...

... a western, proteins are: (1) separated by size, (2) transferred to a membrane, and  (3) detected using...expensive system-specific sandwiches.   Figure 2: The western blot dry apparatus includes a stack of...thiols, to cleave disulfide bonds. Load the samples, one sample per lane, onto the top of a resolving gel...all of the proteins in a sample migrate through the stacking gel at the same pace and enter the resolving...consistent size and density present in all samples. Uneven sample loading or incomplete protein transfer ...Figure 1: Overview of the Western blot process. The sample, a mixture of protein is separated in a gel, transferred...is the first step of a western. To prepare the samples for SDS-PAGE, measure the protein content and normalize...

...another. You’ll need to validate that points 1 and 2 hold true under your specific experimental conditions...when (1) other strategies may not be feasible and (2) large-scale proteomic and transcriptomic datasets...antibody signal in the retinal sample, but little to no signal in the liver sample, which you do (C), so your...epitopes on the same target reduces the likelihood that both antibodies would bind to the same off-target ...to - particularly when you start using complex samples such as whole cell lysate or tissue sections. It...require variable expression of your target across samples - it is hard to distinguish between specific, invariable...there is a wealth of transcriptomic data from this sample type, so you use your bioinformatics skills (or...

...SnapGene Viewer (Mac/PC) FinchTV(Mac/PC) Chromas (PC) 2. You should see individual, sharp and evenly spaced...less and you might suspect contamination in your sample or consider asking your sequencing facility to ...Use a silica spin column for purification of the samples you send for DNA sequencing If your sequencing ...

... fluorescent proteins expressed in the specimen.  2. Fluorescence Activated Cell Sorter (FACs) A machine...photostability of your fluorescent protein when using 2-photon microscopy as this hasn't been well reported...PMID: 18703742. PubMed Central PMCID: PMC2606057. 2. Chtanova, Tatyana, et al. "Dynamics of neutrophil...depth. Two low-energy photons (typically from the same laser) cooperate to cause a higher-energy electronic...

...Transgenesis." PLoS ONE. 8(12): e83043 (20 December 2013). 2. Carola Engler et al. "A One Pot, One Step, Precision...toolkit - aptly named GreenGate – offers all of those same advantages to researchers working in plant model...allows us to design modules that always have the same recognition site but distinct sites where the enzyme...

...linear region to solve for total expression time. 2. Timer functionality with a single gene Although it...290.5496 (2000): 1585-1588. PubMed PMID: 11090358. 2. Tsuboi, Takashi, et al. "Age-dependent preferential...and look at them under a microscope, fractionate samples to isolate particular organelles and their contents...

...;550(1-2):251-268. doi: 10.1016/j.ijpharm.2018.08.043. Epub 2018 Aug 23. PMID: 30145245. Samiul Amin, ...and particle formation. J Pharm Sci. 2015 Feb;104(2):602-11. doi: 10.1002/jps.24259. Epub 2014 Nov 21....

...of these neurons via the channel Channelrhodopsin 2 (ChR2) reduced glutamate transmission from the mPFC...deliver Cre-dependent expression of  Channelrhodopsin 2 (ChR2), a modified receptor protein that activates...actively swimming versus passive floating. In the same paper, the authors used lentivirus to introduce ...

...selection. Desfarges S, Ciuffi A. Viruses. 2010. Jan;2(1):111-30. PubMed . Altering the tropism of lentiviral...Nerhbass U, Montagnier L, Charneau P. Cell. 2000. 101(2): 173-185. PubMed . Woodchuck Hepatitis Virus Posttranscriptional...Lentiviruses can be used to make stable cell lines in the same manner as standard retroviruses. That is, many lentiviral...

...phosphoinositides (Varnai and Balla, 2008) (Figure 2). RNA production can be detected by using a MS2 coat... 296.5569 (2002): 913-916. PubMed PMID: 11988576. 2. Tsien, Roger Y. "The green fluorescent protein." ... proteins are also a better choice for thicker samples. Another solution to avoid autofluorescence is ...reaction and hence, there is no need to excite the sample. Although bioluminescent probes are usually dim...

...rating, sample species, antibody species, or whether the results include knockout data.    Figure 2: Search...

...prompt you to log in or create an account.  Figure 2: The new deposit materials bar New menu Most common...Materials”. The “Search Repository” function is our same search bar. And if you’re in the mood to browse ...

...PMID: 24336571. PubMed Central PMCID: PMC4089965. 2. Sanjana, Neville E., Ophir Shalem, and Feng Zhang...available for a little over a year and a half, a small sampling of the research it has already been used for includes...

...elution choices: TE, Tris buffer, or water Figure 2: Tris-EDTA, Tris, or water? The buffer you choose ... last step will influence the stability of your sample over time and determine which experiments you can... impact the time you will be able to store your sample without having substantial degradation and which... DNA, it will limit what you can do with these samples. This is because divalent cations are essential...reaction, an effective possibility is to dilute your sample in water to reduce the EDTA concentration. Alternatively...

... unknown bacteria. Figure 2: Christina Mork collecting samples at the hot spring. Hiking all...highly complex hot spring sample. The sample was separated into smaller samples of 5-10 cells each for sequencing...leave your samples behind! Paul and Anastasia (Potanina) Neddersen characterized a few samples by 16S ...biological dark matter and so the lab organized a sampling trip to Yellowstone National Park (YNP). Extremophiles...from these extreme environments. Gearing up for sample collection at the hot springs My husband, Paul ...post doc in the Quake Lab, volunteered to do the sampling and I happily agreed to accompany him to YNP the... and two deep cycle batteries for power so the samples could be kept frozen after collection. Figure...

...Information Science and Technology 60.4 (2009): 858-862. 2. Van Noorden, Richard. "Interdisciplinary research...Librarianship and Scholarly Communication 2.2 (2014): 2. Additional Resources on the Addgene Blog 6 Tools...amount of interdisciplinary research while at the same time increasing interdisciplinary citations in their...

...PMID: 25386923. PubMed Central PMCID: PMC4227841. 2. Young, Carissa L., Zachary T. Britton, and Anne S...N- and C-terminal. Each set of plasmids uses the same restriction enzymes for cloning (see schematic below...one need to remove the tag post-purification. A sample of pCri constructs: pCri-1a, -4a and -9a. 6xHis...

...bringing in environmental contaminants.  Figure 2: An Addgenie works in the tissue culture hood while...doi.org/10.1290/1071-2690(2002)038<0086:caeomi>2.0.co;2 Uphoff CC, Drexler HG (2011) Detecting Mycoplasma ...commonly used for every cell culture. Reusing the same bottle of serum again and again for each subculture... changed for at least two or three days before sampling (Uphoff and Drexler, 2011). Culturing on agar...

...from a single Illumina MiSeq® run. As shown in Fig. 2, below, differences in input concentration across ...carrying sample-specific barcodes. Rather than continuing to process 100s to 1000s of samples individually...projects in many labs. However, the high costs of sample preparation for high-throughput next generation...technology overview For any NGS experiment, the DNA sample being sequenced must be sheared into smaller fragments...workflows are limited by costly library prep, as each sample requires an individual library. The plexWell™ workflow...library prep”, in which a relatively large number of samples are subjected to a DNA-barcoding step that uniquely...labels DNA from each well of a 96-well plate. Then, samples from several plates are pooled together to create...