... Commercial Plasmid Prep Kit Amount 1000 Lanes 1000 Lanes 1000 Lanes Cost $200-$500  <$10 $20-30 ...convenient, commercial ladders cost between $200-$500 for 1000 lanes worth of ladder. The Penn State ladders are...plasmid prep protocol, it cost less than $10 to make 1000 lanes worth of the 100 bp or the 1 kb ladder. Using...Your Agarose Gel Other Resources Read about Gel Stains on the BitesizeBio Blog ...

...fight in the Napoleonic Wars and against something even more sinister. 1000 pages long. Incredibly imaginative...larger epic of fantasy novels. Set in a world where 1000 years ago the hero lost and the Dark Lord rules ...founder, Eric Perkins, is Addgene’s Director of Science Product Management and explains the origins of ABC: ... favorite books - both scientific and not - to inspire you to read a little more this October. Science..., examining relationships between microbes and insects, mammals, and plants, among others. I particularly...and with the events of the times."     - Aliyah Weinstein, Marketing and Communications Manager Addgene's... Aliyah Weinstein...

... but a select few are particularly special. Over 1000 of our 60,000+ deposited plasmids have been requested...Said Blue Flame awardee Sophien Kamoun of the Sainsbury Laboratory in Norwich, UK: “We’re very pleased...

...barcodes. Rather than continuing to process 100s to 1000s of samples individually, plexWell™ reduces every...help unlock the potential of modern sequencing instruments by enhancing the efficiency and simplicity of...completely unnecessary for sequencing multi-kb DNA inserts and larger targets. The NGS readout also reveals...

...pieces of roughly equal size, typically between 200-1000 bp. This is to ensure that your downstream analysis... antibody on crosslinked proteins. In the case of histone modifying proteins, which bind quite strongly...that ChIP uses native proteins (so antibodies validated only for denatured proteins may not be a good choice...another. For instance, a sample may need more or less cross-linking to firmly attach the proteins to the DNA...is an extremely useful technique that provides insight into protein:DNA interactions. ChIP works by using...exploring these interactions, and can give valuable insight into DNA expression and protein interactions, including...the downstream analysis process. And ChIP itself begins at...  Crosslinking Once you have your sample, ...

...proximity ligation." Nature methods 3.12 (2006): 995-1000. PubMed PMID: 17072308.  4. Dieck, Tom, et al. "...incorporated into newly synthesized proteins instead of methionine. The labeled proteins can be detected using click-chemistry...turnover of individual proteins in situ.  For the visualization of specific proteins in situ immunocytochemistry...audiences. Right now, she is a science instructor at The Innovation Institute in Boston. When Eugenia is not ...protein is decreased in Alzheimer’s disease patient’s brains. However, it is not known why or how this happens...degradation. These processes control the abundance of proteins in cells. Homeostatic regulation of protein abundance...type. However, we cannot distinguish new from old proteins within that pool. Therefore, it is not possible...

...and very cool 8,768,560 hours – or almost exactly 1000 calendar years!   *Dammit, man, I’m a scientist... to make a plasmid. Doench’s lab in the Broad Institute has created hundreds of thousands of plasmids,...Table 1.   Of course, this is for the simple insertion of a PCR product into a vector. The time it takes...your experiment. Make your work “high-throughput” Instead of making one plasmid at a time, try constructing...

...particles. Studies have found that values can be 10-1000 fold higher than for functional assays depending...The day has arrived; you’ve painstakingly cared for your packaging cell line, prepped your DNA, transfected...

...coefficient and quantum yield of the protein, divided by 1000. In many cases the brightness is compared to that... photoconvertible proteins are PA-GFP, Dendra2, and the mEOS proteins. Some proteins are reversibly switchable...you really know how fluorescent proteins glow?   Fluorescent Proteins (FPs) were first discovered over...categories of fluorescent proteins Photoactivatable / Photoconvertible: These proteins can switch their color... less).  For LSS proteins, the Stokes shift is ≥ 100nm. Specifically, these proteins are excited by UV...when labeling proteins that interact. Indeed, FRET is often used to determine if two proteins interact. ...wavelength than the exciting wavelength. Thus, FPs are proteins with this unique capacity. Many of these FPs are...

...Beroukhim and Boehm labs pulled data from more than 1000 CRISPR screens in DepMap to see if they could identify... for instance, a recent preprint released by the Boehm and Beroukhim labs at the Broad Institute of MIT...MIT and Harvard, Dana-Farber Cancer Institute, and Koch Institute, where an exploration into the effects...introduced an ancestry bias into their experiments instead of a true correlation.   It turned out there was...

... (response elements). Promoters are about 100 to 1000 base pairs long and found upstream of their target...Strong insect promoter from baculovirus Constitutive  Commonly used in expression systems for insect cells...to replicate plasmids and make sure the cells maintains it. Here, we'll cover the promoter, which is necessary...responsible for initiating the transcription of your insert into RNA. In practice, the term "promoter" describes...regulatory factors: enhancers, boundary elements, insulators, and silencers; however, some "leakiness" of ...Strong hybrid mammalian promoter Constitutive  Contains CMV enhancer, chicken beta actin promoter, and...with Tetracyline or its derivatives.  Typically contains a minimal promoter with low basal activity and...

...used to calculate the viral titer: GC/mL = {[(R*C)(1000/V)]*D} R = Reaction volume C = Copies/uL V = Volume...using water-oil emulsion technology. Each droplet contains the ingredients for amplification of the target...experiment. The process to titrate AAV by ddPCR begins with diluting the virus. It is important to note...containing approximately 20,000 droplets. Each droplet contains the material required for a mini amplification...

...of template should have homology arms between 500-1000 bps. HDR efficiency with circular plasmid templates... generally preferred for large insertions such as fluorescent proteins or selection cassettes. This class...conditional and insertion alleles using long ssDNA donors and CRISPR ribonucleoproteins. Quadros RM, et...Stroik January 24, 2023. DNA damage drives genome instability and contributes to cancer, premature aging, and...sticky or blunt ends and often introduces small insertions and deletions along the way. HDR uses homologous...overhang. This will serve as both a substrate for proteins required for strand invasion and a primer for ...template to create mutations HDR donors can contain insertable sequences or specific mutations, along with homologous...

...includes SpCas9, and shortern orthologs approximately 1000 amino acids in size. From the pool of shorter orthologs...split protein like Cas9 is to use split inteins. Split inteins are a pair of naturally occuring polypeptides...toxicity. More AAV-based CRISPR systems Smaller Cas proteins SaCas9 isn't the only CRISPR enzyme that’s small...advantage that it can be turned off by anti-CRISPR proteins. The Sontheimer lab used NmeCas9 with AAV to edit...disease-causing loci in mouse liver.  Two other smaller Cas proteins include Cas12b and CasX. While there are no papers...papers yet published that deliver these Cas proteins with AAV, at just 1,108 and 986 amino acids, respectively...polypeptides that, when at the ends of two proteins, mediates protein trans-splicing, similar to an intron...

...Low 572/ 610 FP595 Tetramer Green Dark/ Red 70 - 1000 Medium 590/ 600   Choosing a photoactivatable...FP-chimeric proteins. Photoactivatable fluorescent proteins (PA-FPs) are fluorescent proteins that display...Fluorescent proteins (FPs) offer scientists a simple yet powerful way to tag cellular proteins and investigate...Threonine 203 to Histidine (T203H) (1). wtGFP normally contains a mixed population of neutral (protonated) and...irreversible PA-FPs are the photoconvertible fluorescent proteins (PC-FPs). PC-FPs can convert from one fluorescent...Selected Irreversible Photoactivatable Fluorescent Proteins Protein Oligomeric State Activating Light ...Selected Reversible Photoactivatable Fluorescent Proteins Protein Oligomeric State Activating Light ...

...design 5 guide RNAs (gRNAs) targeting each of the 1000 genes you’d like to investigate in your next CRISPR...Medicine (CMM), Department of Medicine, Solna, Karolinska Institutet, in Sweden. It can be very time consuming... benefit of working with surface proteins is also that such proteins are easily studied in living cells...Medicine (CMM), Department of Medicine, Solna, Karolinska Institutet, in Sweden. He has a particular interest...screen, each gene is not targeted by only one gRNA, instead many different gRNAs are used for each gene. At...molecules were involved in the phenotype. Surface proteins  My group works on immune cells in different contexts.... Thus, a screen can be used to target surface proteins to determine which contact points are involved...

... E. coli.” ACS Synthetic Biology 4(9) (2015):987-1000. PubMed PMID: 25822415. Find plasmids from this...allowing a spacer-repeat “brick” to be inserted. The BsaI site remains intact, allowing subsequent “bricks...make sure that each cell that gets the plasmid contains all of the desired gRNAs. This increases the chance...the Ventura lab. pX333, a modification of pX330, contains humanized wtCas9 and two U6 promoters. To use ...available from the Bullock lab, and gRNAs can be inserted using Gibson Assembly or SLIC cloning methods....)-containing plasmids. Each destination vector contains GFP, enabling you to select cells with high GFP...enables users to express up to 7 gRNAs. The kit contains different destination vectors depending on the...

...fibroblasts to myoblasts. Cell, 1987. 51(6): p. 987-1000. PubMed PMID: 3690668. 40. Ferber, S., et al., Pancreatic...transdifferentiation of glucagon-producing α-cells into insulin-producing β-like-cells [37, 38]. In 1987, Davis... ES cells and iPS cells into mature pancreatic insulin-producing cells. Cell Res, 2009. 19(4): p. 429-...duodenal homeobox gene 1 induces expression of insulin genes in liver and ameliorates streptozotocin-induced...

...produces point mutations at a rate of up to 1 per 500-1000 bp. In comparison, the DNA replication error rate.... The Moffat laboratory maintains a website for the TKO library that contains up-to-date information and...(3) but its portability to other bacterial strains remains seriously limited, as prior optimizations are...ligase to biotinylate proximate proteins in the cell. These candidate proteins can be enriched via biotin ...of fusion proteins, requires less biotin, and shows enhanced labeling of proximate proteins. Overall BioID2...optimization of industrial yeast strains. However, industrial yeast strains are more difficult to genetically...which “bait” proteins are tagged with Protein A-Renilla luciferase and the “prey” proteins are tagged with...

...Maaike Pols, PhD, a Developmental Editor at F1000Research.  With many changes taking place in the world...journal. An increasing number of funding bodies insist that the research data resulting from their funding...your Addgene plasmid #), and other reagents. F1000Research, among many other journals, has joined a pilot...many journals use to verify whether a manuscript contains all the necessary information to be of maximal...results more valuable. Some journals, including F1000Research, also ask for the raw data behind the figures...several rounds of revision before acceptance. F1000Research uses a novel form of peer review that occurs... Maaike Pols is a Developmental Editor for F1000Research, responsible for assessing the scientific content...