Thestatuses, identities, and demographics of scientists are irrelevant to this great sifting ofvalid versus invalid ideas.

Fulfilling this responsibility, however, is being hindered by a new, alarming clashbetween liberal epistemology and identitybased ideologies.

How did we get here? Science provided solutions to such calamities as famineand plague, transforming them “from incomprehensible and uncontrollable forces ofnature into manageable challenges.”

guides(color=guide_legend(nrow=2, byrow=TRUE))

Don’t waste time and energy collecting metrics you’ll never be able to use. Learn as much as you can from the failures themselves, and then move on.

Many people rush to collect data without first considering whether they can come up with a model powerful enough to interpret the data. I

Analysis of your incident data may shed light on your incidents. But incident data really can’t tell you anything about your reliability.

the diversity of native plants and animals declined with the invasion of alien plants, communities and ecosystems were rarely negatively affected

an alien species is invasive only if it threatens the native biodiversity of recipient ecosystems and negatively affects ecosystem functions, which could lead to detrimental socioeconomic impacts

most known alien microorganisms are pathogens51, their impact is expected to be negative at all levels.

Several invasive microorganisms infect mainly native plants as their primary host

transfer efficiencies between them were similar to those seen with parental partners (0.21)

The resulting constructs (the pMATING series) could be self-transferred through a variety of prokaryotic and eukaryotic recipients employing such a rationally designed conjugal delivery device.

Assuming each sequence in our downloaded database represented a unique species

the antagonistic symbiotic relationship between the wasps and their insect hosts allowed the wasp larvae to acquire insect pathogens that subsequently evolved to benefit the wasp

labs(fill='Legend Title')

Earn 75,000 bonus miles when you spend $4,000 on purchases in the first 3 months from account opening, equal to $750 in travel.

Bins used for plotting:

amperometric sensor was then modified to detect an endocrine disruptor.

optical signals are often difficult to detect in situ5,6,7,8

Some people argue that the Welch’s t-test should be the default choice for comparing the means of two independent groups since it performs better than the Student’s t-test when sample sizes and variances are unequal between groups, and it gives identical results when sample sizes are variances are equal. In practice, when you are comparing the means of two groups it’s unlikely that the standard deviations for each group will be identical. This makes it a good idea to just always use Welch’s t-test, so that you don’t have to make any assumptions about equal variances.

Non-parametric tests don’t make as many assumptions about the data, and are useful when one or more of the common statistical assumptions are violated. However, the inferences they make aren’t as strong as with parametric tests.

In practice, a data set with a sufficient number of replicates forms an approximately normal distribution

Transcription of the araBAD genes under control of the pBAD promoter is induced about 300-fold above a basal uninduced level within 3 s of the addition of arabinose to cells growing on glycerol in minimal salts medium

This system can efficiently create genome modifications in multiple phages without the need for counterselection

Nonparametric statistics are often preferred to parametric tests when the sample size is small and the data are skewed or contain outliers.

when to use parametric versus nonparametric tests [8,9,10]

Noteworthy for its longstanding influence is thebook “Nonparametric statistics for the behavioralsciences” by Siege

Vmax has a low-level natural resistance to kanamycin but is more susceptible to other antibiotics. When selecting on Kan plates, Vmax with KanR will simply grow faster than Vmax without KanR. Larger colonies=KanR. Other antibiotics select against Vmax but must have a dosage change compared to E. coli.

Wash the culture in fresh LB before adding glycerol, to avoid problems with reviving the culture later

ecovery media (5 BHI + v2 + 68 mM Sucrose )

Four inducible promoters were found to function in V. natriegens

Vmax@SGIDNA.com

To prepare V. natriegens cells for −80 °C storage, an overnight culture of V. natriegens was washed in fresh medium before storage in glycerol

For best results, use only LB-Miller agar plates. Do not use LB-Lennox or LB-Luriaplates (Vmax™ growth will be suboptimal).

Vibrio natriegens strain containing a major extracellular nuclease knockout and insertion of an IPTG-inducible T7 RNA polymerase cassette for expression of genes under a T7 promoter.

(a)colE1(b)SC101(c)RSF1010

we successfully demonstrated challenging clonings such as Gibson Assembly with 5 and Aquacloning with 3 fragments as well as 8 part golden-gate reactions.

all downstream manipulations with V. natriegens ATCC14048 Δdns cells were performed with antibiotic selection (kanamycin (200 μg/mL), chloramphenicol (2 μg/mL), carbenicillin (100 μg/mL))

A survey of 230 diverse bacterial and archaeal genomes found evidence of DNA methylation in 93% of genomes, with a diverse array of methylated motifs (834 distinct motifs; average of three motifs per organism)

conjugative plasmids have broad-host ranges23, are resistant to restriction-modification systems24, are easy to engineer with large coding capacities25, and do not require a cellular receptor26 that would provide a facile mechanism for bacterial resistance.

Bacterial cells are typically one thousandth the volume of mammalian cells, which places them near the edge of instrument detection. At this size it can be challenging to differentiate viable cells from debris of similar size

Detailed descriptions, assumptions, limitations and test cases of many popular statistical methods for ecological research can be found in the GUSTAME server (Buttigieg and Ramette, 2014), and in the review by Paliy and Shankar (2016).

condensing the information into two- or three-dimensional spaces. A very good overview of techniques to achieve this was written by Paliy and Shankar (2016).

There are several widely used tool collections, e.g., QIIME 2 [13], mothur [14], usearch [15], and vsearch [16], and 1-stop pipelines, e.g., LotuS [17], with new approaches continually being developed, e.g., OCToPUS [18] and PEMA [19]

Recently, redox-responsive biomolecules such as phenazines have been used in several electrochemical strategies to interrogate a range of biological activities30,31 and to control gene expression in living cells32,33, where the redox status of the biomolecules could be measured or manipulated by application of electronic potentials

There are two main reasons to use logarithmic scales in charts and graphs.

One reason for choosing a dot plot rather than a bar chart is that it is less cluttered. We will be learning other benefits of dot plots in this and future posts.

Using a logarithmic axes on a bar graph rarely make sense.

1000 cells per sample were sorted into microwells by flow cytometry, heat treated to extract DNA, and used as template for a duplex ddPCR reaction

The genre kicked off with “Maps of Time” (2004), by David Christian, and includes such practitioners as Mr. Harari, Steven Pinker, Jared Diamond and Francis Fukuyama.

geNomad uses marker information to demarcate provirus boundaries.

here's a trivial example of why the "standard" geometric mean  is not a good estimate of central tendency when you have values near  zero.

o improve accuracy, Sony hasintroduced the Weighted Least Square Method (WLSM) al-gorithm. In WLSM, the square value of residuals are individu-ally weighted

Impossible Foods, Zymergen, Pivot Bio, Novartis, and Calyxt

and for P. putida, Ap, Sill" Su, or Nal at 200 j.tg/ml, Cm at400 j.tg/ml, and Km or Tc at 25 j.tg/ml

Ramsay, J. P. & Firth, N. Diverse mobilization strategies facilitate transfer of non-conjugative mobile genetic elements. Curr. Opin. Microbiol. 38, 1–9 (2017)

A cryptic rolling-circle plasmid from a commensal Escherichia coli has two inversely oriented oriTs and is mobilised by a B/O plasmid

RIVET has been used as a promoter-trap method to identify bacterial and fungal genes induced during infection (6–8) and as a tool to analyze the spatiotemporal patterns of induction of such genes (9).

Package ‘flowWorkspace’

rbind2(cytoset, cytoset)

taxa should be monophyletic

glyphosate (the most common pesticide used globally) inhibits growth of bacteria

Studying the effects of temperature—along with other climate change-induced environmental disruptions

thepotential "glycosyl transferase" enzymes thathave been chose

docked to theactive site of the glycosyltransferase enzyme by usingAutodock software

However, its hightoxicity restricts its use in the pharmaceutical field

Glycosyltransferaseenzymes catalyse the glycosylation reactions, which leads to a decreasein the toxicity of celastrol.

E. coli BL21(DE3) strain was preferred forthe production of recombinant protein inthis study, thanks to all these properties andits well-known knowledge

This molecule,unlike other VOCs, originated only from lung cancer cells. It has been found that theHeneicosane molecule works in nature as a pheromone of some species.

Celastrol is one of the most important triterpenoid molecules used in ancient times in China

A vector with the mScarlet-I protein construct under the nptII promoter from

conjugation of the TOL plasmid was found to increase with the specific growth rate of both the donor and recipient (Smets et al., 1993, Seoane et al., 2010). This suggests a relationship of faster-growing bacteria (r-strategists) being able to transfer and receive degradative plasmids better than the slower-growing K-strategists

For each culture, a total of 106 cells were added. Naphthalene was added to the NAH7 conjugation cultures by adding naphthalene crystals to the CBM medium which ensured the medium was continually saturated with naphthalene

Novosphingobium aromaticivorans F199donor harboring pNL1; 16S copy number: 3

All data were analyzed using RStudio, version 1.4.1103. Experimental replicates for all data were averaged and reported with standard error.

Bars are colored by whether naphthalene was added and at which hour the sample was taken.

To quantify conjugation via fluorescence-activated cell sorting (FACS), genes encoding constitutive fluorescence of mScarlet-I were inserted onto the NAH7 and pNL1 plasmids.

these transfer regimens were evident for not only the donor transferring the plasmid, but also the recipient receiving it (Sysoeva et al., 2019).

Historically, bacterial evolution has favored two general ecological growth strategies: high reproduction rate (r-strategists or copiotrophs) or optimal resource utilization (K-strategists or oligotrophs)(Atlas and Bartha, 1997)

environmentally-friendly

Care should be taken to avoid severe ecological disturbance when introducing an alien species.

confirmed to participate in protein remineralization in sediments

Abundance and diversity of archaea in coastal ecosystems

Archaea mediate four major processes that strongly impact the coastal blue carbon ecosystems, namely, CO2 fixation, organic biopolymer transformation, CH4 production, and CH4 oxidation

That's greater than taking all the humans who lived throughout time, multiplied by the number of grains of sand on Earth, multiplied by the number of atoms in the universe.

Digestion is recommended whenever DNA input is greater than 75 ng

we show that they have shaped the evolution and host range of plasmids

We found a hierarchy of avoidance, with plasmid genes avoiding 6-bp palindromes significantly more on average than core and non-core chromosomal genes

For the most common target length (6 bp) we show that target avoidance is strongly correlated with the taxonomic distribution of R-M systems and is greater in plasmid genes

As an initial proxy for restriction targets, we first analysed the avoidance of short palindromes in each pangenome component for k=4 and k=6

bacteria possessing cognate R-M systems have higher rates of horizontal gene flow between them

This addictive quality may contribute to their occasional occurrence on MGEs such as plasmids: around 10.5% of plasmids carry R-M systems (Oliveira, Touchon, and Rocha 2014) and experiments have shown R-M system carriage can lead to increased plasmid stability in cells

We report that the EcoRI restriction endonuclease-modification methylase (rm) gene pair stabilizes plasmids that carry it

maths historian George Joseph’s book The Crest of the Peacock: Non-European Roots of Mathematics (1991) and the ongoing encyclopaedic series Science Across Cultures, edited by Helaine Selin

Artificial proteins fine-tuned to five distinct lysozyme families showed similar catalytic efficiencies as natural lysozymes, with sequence identity to natural proteins as low as 31.4%. ProGen is readily adapted to diverse

requires experiments that alter climate in combination with manipulations of microbial communities.

While measurements at each scale do exist for plant-associated microbes, integration across scales is missing for any single system

Data that combine multiplicatively, like rates, are actually very common outside of economics too. The key is to recognize when a measured variables is affected by many (semi) independent forces, each of which scales that variable up or down — rather than simply adding or subtracting a fixed amount to it. This is often true in the natural sciences.

the use of constructed wetlands to generate cellulosic biofuel using waste nitrogen from wastewater treatment

total abundances of archaea and bacteria in these sediments increase with latitude

Increases in solar radiation, temperature and freshwater inputs to surface waters strengthen ocean stratification and consequently reduce transport of nutrients from deep water to surface waters, which reduces primary productivity

received 3-CA every day (press-disturbed), every 2, 3, 4, 5, 6, or 7 days (intermediately-disturbed), or never (undisturbed)

as assessed by 16S rRNA gene terminal restriction fragment length polymorphism (T-RFLP)

Non-metric multidimensional scaling (NMDS, unconstrained ordination) shows temporal dispersion effect

3-CA,41 which is also known to inhibit both organic carbon removal and nitrification in sludge reactors

To investigate their roles, well-replicated time series experiments are needed.

need to understand what governs the relative balance between stochastic and deterministic processes

In a deposition, an engineer at Tesla made this all but explicit: “We want to let the customer know that, No. 1, you should have confidence in your vehicle: Everything is working just as it should. And, secondly, the reason for your accident or reason for your incident always falls back on you.”

The output .clstr file looks like

although the ocean and rainforest seem to be two different extremes of dissimilar environments, surf and turf have several similarities. One similarity that is present in both environments, yet seems counterintuitive is the fact both a reef and a rainforest are essentially nutrient desserts. Both ocean water and forest soil contain low levels of biologically relevant nutrients, and as a result, organisms have developed creative and sometimes symbiotic/mutualistic strategies to thrive in these nutrient-poor environments.

Each flowerpot was filled with soil-bacteria pellets from about 4L of culture, which corresponds to about 20 g (dry weight) of soil

inoculate 50 mL LB in 250 mL Erlenmeyer flasks

At the time of writing, there is an open bug in Snakemake (version 5.8.2) on Windows systems that prevents requesting specific files from the command line when those files are in a subdirectory.

When running on Windows using Git Bash and Anaconda, the previous code will not work. Multiline strings containing multiple shell commands are not executed correctly. The simplest workaround is to add &&\ to the end of all lines except the last inside the multiline shell command:

One thing you’ve probably noticed is that all of our rules are using Python strings

We need to add wordcount.py as a dependency of each of our data files so that the rules will be executed if the script changes

Before sequencing your plasmids, we linearize them so that we get mostly full-length sequence reads

If the pipeline does not produce a consensus for your target, you can download the raw reads from your dashboard and bin them yourself, but please note that raw reads are much more noisy and error-prone (~98.3% accurate) than consensus reads.

The SWW aimed to mimic the average conditions and nutrient proportions of conventional domestic wastewater.

However, long-term live imaging is laborious and equipment intensive, because a single microscope often has to be monopolized for the duration of the experiment

For each of the libraries, we screened for the presence of all possible STMs used in a given experiment, with the requirement that all spike-ins for a given STM needed to be present

The advantage of merging these paired-end reads later is that the quality can be improved (if overlapping bases are the same in both reads, there is a higher likelihood that the correct base was called) so there is no need for trimming here.

In the absence of inducer, basal activity of these EilR-regulated promoters fell below that of the three routinely used inducible systems PBAD, Ptet, and Ptrc

We estimated read counts for eachisolate by assigning trimmed and merged reads that perfectly matched a known isolate 16S rRNAV4-V5 sequence to that isolate. Reads that did not match a known isolate were discarded.

that if mutations are sufficiently abundant (for instance, if they are generated at an early PCR cycle), they might still give rise to a spurious OTU.

a flavour with the same Ubuntu engine under the hood, but with a lightweight desktop environment, and with lighter application programs. the ultra-light Lubuntu or the medium light Ubuntu MATE or Xubuntu.

Primers to target the DXS gene24 (one copy per genome) and the ampicillin resistance gene on the plasmid were created using the Primer3Plus design tool

With this simple realization, a number of groups have realized that HTS datasets are actually count-compositions

Identification of type IV conjugative systems that are systematically excluded from metagenomic bins

the box plot of ANCOM-BC had the shortest width, suggesting that it not only successfully estimates the true sampling fractions and eliminates bias due to its variability, but it also has the smallest variance which is not the case with other methods.

According to an extensive simulation study1, among the available methods for DA analysis, only ANCOM performs well in controlling FDR while maintaining high power, as long as the sample size is not too small

green fluorescent protein containing a degradation tag (GFPmut3_LAA)

Consequently, amplicons produced with these primers were removed leaving amplicons corresponding to eight variable regions (V1, V2, V3, V4, V5, V6, V7, and V8) for the MVRSION method.

DMSO has been successfully used to improve the performance of RT-PCR [10] and SangerSequencing [11].

This consensus .fastq file will be delivered with your standard sequencing results, whereas the .fastq file for the raw read data must be requested during order submission (as above) and will be delivered via a separate raw data link.

As such we believe that it was important to test these methods on a wide range of different real-world datasets in order to gain an understanding of how they differed from one another

If every number between 0 and 1 had equal probability of being chosen, method 2 would be a valid hypothesis test—but not a good one.

The advantages of using SSU rRNA for community fingerprinting are many: (i) This gene is found in all cellular life forms. (ii) It is a highly conserved gene, serving to a large degree as a reliable molecular chronometer. (iii) It is seldom transferred horizontally. (iv) It possesses both conserved and variable regions, so that the conserved regions can be targeted by polymerase chain reaction (PCR) primers and the variable ones be used as identifying markers. A handful of other genes, such as the large subunit (LSU) rRNA share these properties, but the length of ~1,500 bp of the bacterial SSU rRNA made it amenable to early molecular techniques, and the impressive body of knowledge that has since accumulated on the basis of this gene makes a switch to other markers very impractical, except in certain sub-fields such as mycology, where ITS and LSU are widely used.

QuantStudio™ 7 Flex Real-Time PCR System features a decoupled six-by-six-color filter set thatsupports the dyes shown in the following table.

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Path/to/AppImage

chmod +x *.AppImage

Fundamentally, all real-time thermal cyclers are engineered to detect fluorescein (FAM) first and foremost, so dyes with longer wavelength emission may be detected less sensitively

For those shorter than 25 bases, a standard (dual labeled) BHQ™ Probe may be perfectly satisfactory.

the majority of chimeras are believed to arise from incomplete extension. During subsequent cycles of PCR, a partially extended strand can bind to a template derived from a different but similar sequence. This then acts as a primer that is extended to form a chimeric sequence

Koodankulam: What the Media Didn’t Tell Us

Rahul Gandhi’s comparison is great fodder for intellectual debate, but it is not good politics. When the country is in ‘wartime’, the adornments of peace do not sit very well. This comparison is a peacetime jewel. In a war zone, only the cannons, pistols and swords do well.

To perform bacterial genome assembly, we suggest using the third-party de novo assembly tool Flye3. This analysis package represents a complete pipeline, taking raw nanopore reads as input, and producing polished contigs as output. We also recommend one round of polishing with Medaka4. These tools can be found on GitHub

droplet digital PCR for simultaneous detection of plasmid and genomic DNA in sorted cells

For each of these two subpopulations, 1000 cells were sorted

pop.MFI computes and returns the median fluorescence intensity for each marker. They are typ-ically used as the arguments passed to gh_pop_get_stats method to perform the sample-wisepopulation stats calculations

Additionally, make sure to use both forward and side scatter on log scale when measuring microparticles or microbiological samples like bacteria. These types of particles generate dim scatter signals that are close to the cytometer’s noise, so it’s often necessary to visualize signal on a log scale in order to separate the signal from scatter noise.

By introducing rationally selected combinations of folding-enhancing mutations into GFP templates and screening for brightness and expression rate in human cells, we developed mGreenLantern, a fluorescent protein having up to sixfold greater brightness in cells than EGFP

But Poisson statisticspredicts that there will be some empty droplets (1,642 empty droplets) and gives a preciserelationship between the average number of copies/droplet and the expected fraction ofempty droplets.

Note that for creating perfect PDF files for printing, it’s better to turn to a dedicated desktop publishing software such as Scribus, which can also import SVG files.

10-20 sec

3 uL diluted ROX per 20 uL reaction