...initiate replication, controlling the amount of active DnaA available is one of a cell’s best tools to...binding sites, or trading out ADP for ATP to re-activate DnaA. These regulatory mechanisms allow E. coli...machinery in order to replicate—but not always to the same extent. As described in our previous Origin of Replication... although ATP does get hydrolyzed over time, inactivating the protein binding it (Grimwade et al., 2018...proteins, it can be regulated indirectly via the same mechanisms that affect DnaA and IHF. RepA also controls...

...Golden Gate assembly involves the simultaneous activities of a Type IIS restriction enzyme such as BsaI...base pair substrates, it demonstrates significant activity on some mismatch-containing pairings. During Golden...confirmed the insertion of the Lambda insert at the same high frequencies as the antibiotic selection markers...

...efficient polymerase activity, whereas some ratios result in significantly reduced activity (4). To ensure ...technically possible to encode all four viral genes in the same plasmid (as opposed to four separate plasmids), ...

... enzymes are cheap, fast, shelf-stable before activation, and light-insensitive. They can detect femtogram...usually preferred because of their higher specific activity, slightly lower cost, and smaller size. However...antimicrobial agents. Chemiluminescence kits When activating your enzyme, make sure to choose a chemilumescence...and your sample(s), as you can only compare samples to a standard curve generated on the same blot. If...protein expression varies from sample to sample. To compare one sample to another, you’ll need to normalize... a weak signal. However, they have high cross-reactivity, vary significantly from lot-to-lot, and have...but they have higher specificity, lower cross-reactivity, and are more consistent lot-to-lot. The highest...

...going forward, Seeding Labs hopes to expand its activities by facilitating connections to mentorship and...lines of emerging global challenges. However, these same researchers face significant barriers to scientific...

...control recombinase activity. For example, post-transcriptional recombinase activation via RNA toehold riboregulation...states in a sample; instead, it will suggest the predominant state of registers in a sample based on a ... use the same recognition site array on the register, and all three-input GRSMs use the same recognition...implement the same gene regulation program. We recommend testing multiple register designs for the same gene regulation...If an attB-attP pair is aligned (sites facing the same direction) on a register, then the result of their...amplify in some states but not others. Because samples of many registers will likely not be homogenous...infer the predominant state of registers within a sample using a metric that we describe in the Materials...

...in our hands, is both cost-effective and highly active. After csRNP complex formation, a single-stranded...efficiently and serves as a good control for both the activity of CRISPR reagents and ICC antibody specificity...of the crRNA (aim for 40-60% GC content) crRNA activity Local chromatin structure (although we could successfully...turnaround time 5-7 days); because the tracrRNA is the same in all csRNP experiments, it can be ordered in bulk...

...new deposit materials bar New menu Most common activities can be done straight from the new search bar,...Materials”. The “Search Repository” function is our same search bar. And if you’re in the mood to browse ...

...transcriptional repression, activity dependent de-repression, chromatin remodeling, gene activation, and more. Loss...gene replacement therapy, gene editing, MECP2 reactivation, RNA editing, RNA trans-splicing, and protein...They introduced a mouse MECP2 guide RNA and a hyperactive version of the human RNA base editing enzyme ...https://doi.org/10.3389/fgene.2019.00625 McGraw CM, Samaco RC, Zoghbi HY (2011) Adult Neural Function Requires...

...are insoluble, act on DNA, or are enzymatically active. For this reason, recombinant proteins are typically...* Streptomycin and Tetracycline Contains highly active thioredoxin reductase and glutathione reductase...proteins. hsdSB (rB- mB-): These strains have an inactivated native restriction/methylation system. This means...-deficient; allows for cloning and expression in same strain F- recA1 hsdR(rK12- mK12+) (DE3) (RifR) ...plasmids utilize inducible promoters, which are 'inactive' until an inducer such as IPTG is added to the...

...purification can be one of the most stressful lab activities. Working with proteins requires a substantial...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...

...have allowed for real-time monitoring of neuronal activity across substantial 3D volumes. In addition to ...Weijian, and Rafael Yuste. In vivo imaging of neural activity. Nature Methods 14.4 (2017): 349-359. PubMed PMID...due to the refractive index mismatch between the glass and the water-like buffer the sample is immersed...microscope to use for a new sample I always start by asking two questions: 1) Is the sample dynamic or static? ...? 2) Is the sample thin (< 15 μm) or thick? These questions will place each sample into one of four categories...capture in these types of samples is their fast movements. Depending on the sample, a temporal resolution... fluorophores higher in the sample. TIRF is also very gentle on the sample as the majority of the laser...

...genome to be probed for proteins with host-binding activity without the need for genome sequencing or annotation...nitrocellulose for subsequent probing for cell-binding activity.  First, cut out 20 Petri dish-sized circles of...at the same location that their host colonies grew. If your cells of interest stick to the same location...

...population in a procedure termed fluorescence activated cell sorting or FACS (2). By analyzing single ..., Sweet RG, Herzenberg LA (1972) Fluorescence Activated Cell Sorting. Review of Scientific Instruments...was raised in the same host species, of the same Ig subclass, and conjugated to the same fluorochrome as...ensure that samples are single cell suspensions. For cells that tend to aggregate, pass the sample through...using an amine reactive dye, such as LIVE/DEAD stain. Like DNA binding dyes, amine reactive dyes can only...single-stained samples for each fluorochrome in the experiment. Single-stained samples must be as bright...experimental sample. In addition, you should include a panel of “fluorescence-minus one” controls, samples stained...

...  LacZ β-galactosidase activity abolished   lacY Lactose permease inactivated, lactose cannot be taken...increases plasmid stability recBCD Exonuclease V activity abolished Increased plasmid stability, reduced...convoluted due to researchers sharing and renaming samples throughout history. It was likely isolated in 1918...taken up by cell mcrA, mcrBC Inactivation of pathway that cleaves methylated cytosine DNA Allows for ... foreign (methylated) DNA mrr, Δ(mcrC-mrr) Inactivation of pathway that cleaves methylated adenine or...source leuB β-isopropyl malate dehydrogenase inactivated Requires exogenous leucine source for growth ...proAB lacIqZΔM15 Tn10(TetR Amy CmR)]  Note: Inactivating mutations in specific genes are signified with...

...chemogenetics (Designer Receptors Exclusively Activated By Designer Drugs, DREADDs) and optogenetics as...use of DREADDs (Designer Receptors Exclusively Activated By Designer Drugs) to determine the role of serotonin...chemogenetic receptors delivered by AAV within the same cell types. Not only can we compare these methods...finger nucleases, and many others (12). The most attractive potential of AAVs is their ability to be used...McCarty, D. M., Paul E. Monahan, and Richard J. Samulski. "Self-complementary recombinant adeno-associated...PMC190186. 9. Grieger, Joshua C., and Richard J. Samulski. "Packaging capacity of adeno-associated virus...

...enzyme that has been mutated to block its catalytic activity and instead become trapped in a covalent bond ...brightness, stability, or other properties is an active area of research. If size is a concern for your...sequentially adding different ligands to the same sample to label sub-populations of your protein of interest...probably find lots of different plasmids containing the same protein of interest tagged with different fusion...affinity purification tags, another version with that same red fluorescent protein somebody made because they...ligands (Gautier et al., 2008). Although based on the same original enzyme, SNAP-tag and CLIP-tag show good...specificity for their own ligands with minimal cross-reactivity. Other strategies include using engineered ligases...

...G., et al. "Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9...targeting the human kinome. These plasmids contain the same 3-4 gRNAs found in the first subset of the pooled...

...proteins in tissues, cells, and biofluids. Recently, Sam Myers, a Scientist in the group, developed a CRISPR-based...and RNA sequencing of complex heterogeneous RNA samples. Furthermore, such high processivity enzymes and...specific genomic loci. He made use of catalytically inactive dCas9 by fusing it to an engineered peroxidase...Using these lentiviral plasmids, catalytically inactive Cas9 (dSpyCas9) fused to APEX2 is expressed from...

...A) Betacyanin is produced from tyrosine by the activity of 3 enzymes and a spontaneous step. (B) RUBY ...GUS staining or are not suited to studying large samples. RUBY Red But recently, something new has been ...consisting of these three enzymes expressed under the same promoter with self-cleaving 2A site linkages and...transformation next to an untransformed seed from the same plant beginning to germinate. (Top right, second...