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We narrowed to 419 results for: SAM-2

Showing: 221 - 240 of 419 results
  1. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ...retain their individual activities when separated (2), and that DBD and TAs from different systems could... Enzymol. 1983;100:293-308. PubMed PMID: 6312261. 2. Keegan, L., Gill, G., and Ptashne, M. Separation ...libraries of bait and prey against each other at the same time in order to try and access the entire interactome...the protein partner or the UAS/reporter gene, the same bait and prey libraries can be screened using both...
  2. New Tools Enable CRISPRa for Neuroscience Applications

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    Blog Post
    ...with neuron-specific expression of dCas9-VPR (Figure 2), a popular Cas9 activator developed by the Church...ubiquitous ones such as EF-1α and CAG. Figure 2: Dual lentivirus system with ubiquitous expression... Gene isoforms refer to mRNAs generated from the same gene but with different untranslated or coding regions...transcriptional start sites, all of which yield the same BDNF protein. To demonstrate the specificity that...
  3. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

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    Blog Post
    ... pCRISPomyces-2 contains Cas9 and a gRNA cassette. The simpler system of pCRISPomyces-2 displays a higher...and a subsequent curing protocol that requires only 2-3 hours incubation. E. coli and T. citrea scarless...four major components including (1) a CRISPR RNA, (2) four proteins forming the QCascade DNA-targeting ...https://doi.org/10.1038/nbt.3043 Bikard D, Jiang W, Samai P, Hochschild A, Zhang F, Marraffini LA (2013) Programmable...
  4. pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs

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    Blog Post
    ...blood are reprogrammed immediately after collection [2]. PBMCs display unique epigenetic signatures that ...PMID: 27882265. PubMed Central PMCID: PMC5118044. 2. Kim, Y., et al., The Generation of Human Induced ...blood sample, in the best-case scenario [7]. In the clinical setting where patient blood samples are limited...grade hiPSCs from easily accessible patient blood samples. All pCXLE vectors listed below in Table 1 are ...
  5. Advice on Career Paths and the Green Card Process for International Researchers and Entrepreneurs

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    Blog Post
    ... both substantial merit and national importance; (2) The foreign national must be well positioned to advance...outstanding achievements in your specific academic field, (2) that you have three 3 years of research experience..., Casseus Law suggests a mix of experts including 2 dependents –people who have worked with you i.e. your...working in October of the same year. The Industry H-1B visa is not the same as an academic H-1B visa and...individually, as a judge of the work of others in the same or allied academic field Evidence of original scientific...
  6. 6 Tips for Grant Writing

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    Blog Post
    ...days before the deadline isn’t going to be much use. 2. Originality is essential for grant success The two... it’s important not to throw in the towel if 1 or 2 funding bodies say no. If possible, get feedback from...institution). Likewise, if you’re staying in the same lab where you have been working for the last few...
  7. Savvy Advocates Needed to Navigate a Scientific Enterprise in Flux

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    Blog Post
    ...is funded have been well documented over the past 2 years (Alberts et al, PNAS 2014; Taylor, eLife, 2016...PMID: 24733905. Pubmed Central PMCID: PMC4000813. 2. Taylor, Justin Q., et al. “Avoiding a lost generation...postdocs at institutions, will be challenging. At the same time, there is much opportunity to make headway ...
  8. Antibodies 101: Beyond Surface Labeling

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    Blog Post
    ...everything but the CD8+ T cells (Figure 2). Even better, we can use the same fluorophore for the three lineage...     Figure 2: Dump gating labels all unwanted cell lineages with the same fluorophore to easily...FACS achieves greater purity of your sorted cell sample than MACS. An effective strategy to sort out your...different while the permeabilization solution can be the same). Examples of such targets include transcription...undesired cell populations. Dump gating uses the same color for different markers, making it easy to gate...
  9. Plasmids 101: Degron Tags

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    Blog Post
    ...required components are 1) a SMASh-tagged protein and 2) asunaprevir (small molecule). dTAG The degradation...gone through an additional round of evolution (AID-2) to reduce some of the leakiness associated with the...the general concept of the system has remained the same. An obvious drawback of the system is that it requires...
  10. Google Forums Round Up: First Impressions of NgAgo

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    Blog Post
    ...engineering methods have their own pros and cons (2). The NgAgo DNA guide is inexpensive and can be 5’...Nature biotechnology (2016). PubMed PMID: 27136078. 2. Burgess, Shawn., et al. "Questions about NgAgo." ... to deliver NgAgo ingredients into the cell, the same method used by Gao et al, 2016 (1). View the Complete...
  11. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

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    Blog Post
    ...from GFP and the mutation definitions found in Table 2 may not apply. Table 1: The specific mutations comprising..., V163A EBFP F64L, S65T, Y66H, Y145F   Table 2: Functional role of specific mutations in GFP derivatives...that of the gene of interest, or expressed from the same transcript as the gene of interest, but after an...
  12. Plasmids 101: Stringent Regulation of Replication

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    Blog Post
    ...the chromosome, ending replication.   Figure 2: Prokaryotic chromosomal replication. Adapted from...Microbiology and Molecular Biology Reviews: MMBR, 62(2), 434–464. https://doi.org/10.1128/MMBR.62.2.434-464.1998...machinery in order to replicate—but not always to the same extent. As described in our previous Origin of Replication...proteins, it can be regulated indirectly via the same mechanisms that affect DnaA and IHF. RepA also controls...
  13. CRISPR 101: Validating Your Genome Edit

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    Blog Post
    ...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...
  14. Antibodies 101: The Basics of Western Blotting

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    Blog Post
    ... 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...
  15. Antibodies 101: Validation

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    Blog Post
    ...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...
  16. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

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    Blog Post
    ... 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...
  17. 6 Tips for Analyzing and Troubleshooting Sanger Sequencing Results

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    Blog Post
    ...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 ...
  18. Quick, Versatile Plant Transgenesis with GreenGate Plasmids

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    ...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...
  19. Fluorescent Proteins 101: Fluorescent Protein Timers

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    Blog Post
    ...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...
Showing: 221 - 240 of 419 results