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...normal' sample with known copy number set as the expression baseline. Copy number is then determined by comparing...calculations so that your assay output number (RQ value) matches the copy number of your sample. In that case, ...comparing the number of positive droplets for the gene of interest and reference gene, copy number can be determined...run, as the method is dependent on number of samples rather than number of cycles.  Choosing a method  As...exact copy number and it may not be able to identify trends in cases where the copy number variation is... to quantify said copies. They do so using copy number variation (CNV) assays. There are several varieties...Quantitative/real-time PCR (qPCR) has been used for copy number assays for years. qPCR works like regular PCR, ...

...re using a PO number. Follow these steps to place an order online while using a PO number: Log in to your...invoice will reference your Addgene order number and PO number that you entered during checkout. At this...does a customer place an Addgene order using a PO number? Why do I have to order directly from your website... website, and you will be able to enter your PO number as payment during checkout. Placing your order ...addresses provided. How do I place an order using a PO number? Placing an order on Addgene’s website is quite...option to “Pay by Purchase Order.” Enter the PO number you generated through your organization in this...Center article How do I pay by purchase order (PO) number? You can find answers to other ordering questions...

...plasmid copy number in a cell. One can attempt to eliminate fluctuations due to copy number by introducing...plasmid copy number is the input and gene expression is the output. The input, plasmid copy number, directly...expression due to copy number is kept in check. In such an iFFL, as plasmid copy number increases, the amount...expression and plasmid number. In a suite of plasmids modified from pSC101 with copy numbers ranging from 3-100...growth stage can both affect gene copy number. Moreover, gene copy number is affected differently by growth... plasmid copy number. During the shift from exponential to stationary phase, the number of plasmids per...specific purposes. Unfortunately, plasmid copy numbers vary within cell populations and over time resulting...

...2001). The number of iterons encoded on a given plasmid influences how high the copy number needs to get... In general, these plasmids tend to be low copy number. Conversely, plasmids that can initiate DNA replication...termination. These types of plasmids tend to be high copy number. Here, we will consider the replication mechanism...initiation of replication. Once the average copy number for a given plasmid has been reached, negative ...replication of the plasmid to balance the total plasmid number.  Let’s consider pSC101. This plasmid is stringently...stringently controlled and has a low copy number. Like the E. coli chromosome, it has a single ori and requires...affect DnaA and IHF. RepA also controls the copy number of pSC101 through a negative feedback cycle. As...

...greater influence, the average number of citations per year and the number of other manuscripts that heavily...the number of citations that paper has is displayed at the top of the page, as well as the number of citations...is measured by the number of citations and the context of these citations. The number of manuscripts that...publications by an author, total number of citations, and the number of publications that heavily cite... Now, search results display a bar graph of the number of papers with your search term published each ...to easily access it later. Pro tip Checking the number of retweets, likes, and comments on a tweet about...to focus your search results. A bar graph of the number of papers with your key term published each year...

...cumulative numbers of CRISPR plasmid distribution and deposits (Figures 1A and 1B); the cumulative number of ...Cumulative number of deposited CRISPR items, categorized by CRISPR function, by year. (B) Number of requests...CRISPR plasmids (Figure 1C), and the cumulative number of CRISPR libraries Addgene has distributed (1C... deposited with Addgene by year. (C) Cumulative number of laboratories that have deposited CRISPR plasmids...categories (Figure 2C). Note that in Figure 2, the numbers on the X axis refer to distribution and deposits...using locally estimated scatterplot smoothing. (C) Number of requests per year for pooled libraries, categorized... made available through the repository. And the number of plasmids and libraries deposited and requested...

...copy number  Although the sequence and regulation of the ori dramatically affect the copy number of a...RNA or proteins, respectively. A plasmid's copy number has to do with the balance between positive and... in mind that plasmids with low to medium copy numbers can still express massive amounts of protein given...below for more details.   Common Vectors Copy Number+ ori Incompatibility Group Control pUC ~500-700...table defines common cloning vectors, their copy number, ori, and incompatibility groups. Note the A -C...group should not be co-transformed. +Actual copy number varies. See below for additional considerations...volume, antibiotic, and medium can all affect copy number. Some ori's are temperature sensitive; others ori's...

... more plasmid. Plasmids can vary in copy number, the number of plasmid copies supported within each bacterial... bacterial cell in your culture. Copy number is partially specified by a plasmid’s origin of replication...inoculation. To further complicate matters, plasmid copy numbers are often ambiguous and should be viewed more as...guideline, since they are typically based on the copy number of the empty plasmid backbone or a researcher’s...experiencing low yields from a plasmid whose copy number is unknown, it is entirely possible that you are...'ve reached your lysate volume limits, the copy number of some origins of replication (e.g. p15A, ColE1...optical density of the culture can indicate the number of cells present in the given volume.  Let oxygen...

...transduction, usually without any numbers for context. That’s because the number can vary depending on your ... enters the cell. Different serotypes utilize a number of different receptors, co-receptors, and other... the most flexibility for expression in a large number of cell types and are useful if you're going to... different systems or want to transduce a large number of different cells. Promoters such as chicken-beta-actin...define high titer, then, is this: It’s the optimal number of viral particles per volume required to express...adequate level for your experimental measurements. The number itself is determined through validation and optimization...strongly recommended to optimize to the “lowest number required” instead of using the first amount that...

... to reduce the number of entrants into PhD programs to more closely reflect the number of positions available...and functions” “Complete lack of information on number of postdocs” “Postdocs are really hired to produce...collection still remain: What are the current numbers of trainees? Are there too few or too many at different... Are there other solutions, such as increasing numbers in Master’s level programs? What would be the effects...College Park, is a strong supporter of increasing the number of postdoctoral scientists at University of Maryland...published an article arguing against reducing the number of graduate students accepted in STEM fields (6...

...replication when copy number in the cell is high, but allows replication when copy number in the cell falls...replication machinery (del Solar, 1998). As the copy number of the plasmid increases, increased amounts of ...promoter and inhibit its own transcription. As copy number increases, RepA transcription is repressed and ... blocks replication machinery when plasmid copy number is high (del Solar et al., 1998).  In cases of ..., so each plasmid “thinks” it has a higher copy number than it actually does, and neither plasmid is maintained...mechanism of incompatibility mediated by low-copy number plasmids. Plasmid 80:54–62 . https://doi.org/10.1016...

... verify a number of key properties (brightness, photostability, oligomerization) of a number of promising...can be quantified for individual cells, and the number of 'normal' looking cells can be counted. This ... we provide an example of the OSER assay with a number of fluorescent proteins. Fusions of CytERM with...Many fluorescent proteins are available, and their number is steadily increasing. Of course, you want the...publications are only characterized under a limited number of conditions, often using purified proteins. Therefore...

...into additional plasmids thereby increasing the number of recombination events that are required to generate... the volume of the reaction, and the timing and number of harvests will all impact the titer. Importantly...virus, consider scaling up by either increasing the number of 10 cm2 dishes or increasing the vessel size....flask will require some initial optimization; cell number, media volume, and plasmid concentrations will ...vessel to the 10 cm2 dish and adjusting the cell number, plasmid concentration, and experimental volumes...routinely, you may opt for simply increasing the number of 10 cm2 dishes. While this option is attractive...

...the assembly (number of inserts or modules) and the resulting efficiency of assembly (number of transformants...transformants); the greater the number of inserts, the lower the number of transformants. To compensate for ...These improvements (5- to 12-fold increases in the number of transformants while maintaining high levels ...overhangs for any Golden Gate assembly design. Number of fragments assembled Golden Gate Assembly protocol...

...power in numbers! When starting off with a new cell line, grow the cells, aliquot them into a number of vials...samples Keep track of passage numbers Keep an eye on the passage number. Cells change their growth rate...rate and morphology at high passage numbers (>30-40 times), depending on the cell line so old cells should...date, the cell line you’re using and its passage number. Especially in a lab where multiple people are ...

... search is... The Addgene plasmid number: This is the unique number given to every plasmid in our catalog...catalog - if you know the catalog number of the plasmid you are searching for, this is the quickest and ...your colleague’s plasmid. Use the Addgene catalog number to easily find template citations for Addgene plasmids...Addgene, you can list the original vector’s plasmid number here. Gene/insert: This describes what has been...bacterial strain in which the plasmid is shipped. Copy number (if known), antibiotic resistance, and optimal ...

... barcoded with a unique ID number (1D barcode as shown below). This number ensures that every sample can... deposits is received in the lab, although this number fluctuates throughout the year (see graph below... to 96 tubes (see image below). Each plate has numbers and letters to give coordinates, from A1 through...each plasmid so that we never run out. There are a number of different strains of E. coli that can be used...ID’s are unique, consistently labeled, and that numbers are not switched around or misread.  2. Track the...

...the key figures Number of requests per reagents (as a horizontal bar chart) Number of requests per ...valuable insight to the most requested plasmid, the number of requested plasmids, or trends of requests over...per country (as dot plot) Number of requests per year (as a bar chart) A written summary of the data,...of dashboards are (i) easy access to important numbers and trends, (ii) user interaction with the data...

...request that you cite it using both its Addgene ID number as well as the reference for its original publication...plasmid from Addgene using its unique Addgene ID number, your publication will be listed on our website...-  While it is tempting to rely on the citation number when comparing two plasmids, we recommend reviewing...can search for plasmids by name, PI, Addgene ID number, or keyword. Although an initial search might give...in the repository, our scientists have curated a number of special collections pages. These pages serve...

...spinning disk microscopy, a metal disk containing a number of holes rotates through the excitation light path...axial volume of a sample. Therefore, when a large number of optical slices are acquired all fluorophores...in the final image. Over the past two decades a number of techniques have been developed that allow for...example is the lattice lightsheet that projects a number of thin cone-shaped beams into the sample. These...require optical sectioning, but open the door to a number of other techniques that can improve contrast and...when imaged. Super-resolution microscopy uses a number of optical and chemical ‘tricks’ to turn subsets...