Microsoft word - 06 app note automated colony counting.doc

Automated colony counting for molecular biologists
How it can improve productivity when selecting recombinant clone
Transformation efficiency is a measure of the amount
transmitted and darkfield illumination, enabling colonies of cells within the baceterial culture that are able to take and plaques to be counted on a range of media types.
up DNA molecules. Some applications such as theconstruction of genomic libraries require that the With åCOLyte Click’n’Count, enumeration is at least as fast as any light box counter because counting involves Counting of recombinant colonies is one criteria used to pointing at each colony and clicking to count. However, calculate transformation efficiency.
since the results are automatically transferred to PC,the time consuming data keying step is eliminated. For A common method for selecting recombinants involves researchers wanting to increase sample throughput, altering the antibiotic resistance, eg, resistance to Synbiosis offers a fully automated version of åCOLyte, ampicillin or tetracycline, cells that contain the vector SuperCount. The system can count up to 103 colonies (plasmid) will survive and cells which do not contain the in less than two seconds, while also automatically vector (plasmid) will not survive. Another method is to correcting for background variations and different use a vector (plasmid) such as pUC which has a LacZ media types and can provide time savings of up to 90 gene present. By inserting DNA within the LacZ gene per cent on a manual count. This greatly increased this will inactivate this gene and colonies will remain laboratory productivity allows molecular biologists to white. Plasmids which do not contain the inserted DNA make instant decisions about whether to discard plates will have an active LacZ gene present producing blue or continue working with the recombinants they have.
For molecular biologists that have plates where the Manual counting of recombinant clones – the
colony counting is complicated by having for example drawbacks
touching or non-recombinant satellite colonies present, Manual methods of enumeration require users to count Synbiosis recommends the ProtoCOL systems. These colonies or plaques using a light box and pen and then are fully automated colony counters, which integrate a transfer the results into a computer. This is not only a CCD camera with extremely sophisticated software and time consuming and tedious task but can leave the way each comes complete with its built-in own PC. The open for misinterpretation with plate reading (especially software features a unique auto-separation algorithm, which means touching colonies can be automatically, colonies). Computer keying errors can also occur. In enumerated as separate ones, thus reducing the need computerised image of the plate alongside the count experiencing problems with small non-recombinant there is no means of carrying out an independent audit satellite colonies, the software allows differentiation of the results to check for accuracy.
based on definable size limits so small colonies can beexcluded from the final count. In addition, because the Automated
software allows users to set the size of the round frame recombinants
placed on the plate image, it means that edge effects such as bubbles or pen markings can also be resistance involves a total count of all colonies or eliminated. Other problem areas within the plate, for plaques on the plate because only those recombinants instance, agar lumps or fungal contamination can also that have antibiotic resistance will grow. Synbiosis be removed from the count by manually drawing recommends the åCOLyte or ProtoCOL systems for around them on screen. All these features ensure the any types of total count because both systems can count colonies and plaques on spiral, pour and spreadplates.
Both the åCOLyte and ProtoCOL systems produce live,full-colour on-screen images that can be saved with a For molecular biologists with a limited budget that have time and date for GLP compliance. Images can be to perform very straightforward counts of recombinant archived for future reference, if necessary, or can be colonies or plaques, åCOLyte is the perfect choice.
printed out for reports or presentation material. This åCOLyte is a cost-effective, yet accurate alternative to feature is important to companies involved in using using a traditional light box and pen.
recombinants for drug discovery research because itprovides secure records that are compliant with the The system comes complete with software, which can information required by external regulatory auditors.
be quickly installed on virtually any laboratory PC. Italso has two arrays of LEDs allowing incident,
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Application Note 6
different colour colonies
Differentiating and counting of recombinants based on
their colour are more demanding tasks to perform
manually. This is because they rely on molecular
biologists being able to accurately define by eye
different coloured colonies, which in the case of the
blue white selection system can vary from one
researcher to another. To overcome this subjectivity in
manual colour differentiation, Synbiosis recommends
the colour option for its ProtoCOL systems. This
enables automatic discrimination and counting of, for
example, blue and white colonies on the same plate.
This means the colour discrimination can automatically
be set at an agreed level by all the users in the
laboratory thus eliminating the need to spend time and
effort deciding if a colony is white or coloured before
counting it.
discrimination application is set at the correct sensitivityfor Conclusions
The flexibility of the åCOLyte and ProtoCOL systems
means that they can be used to rapidly count antibiotic
resistant recombinant colonies on spread or spiral
plates, as well as colonies and plaques on pour plates.
The åCOLyte is excellent for simple total counts,
whereas when the count is more complicated with
different sized or touching colonies involved the
sophistication of being able to discriminate and countrecombinant plaques or colonies of different coloursfrom the same plate, Synbiosis’s ProtoCOL with thecolour option is ultimately the superior system to use.
In summary, the use of automated colony counting forenumerating recombinants can eliminate a substantialpart of an unpopular task, as well as take a fraction ofthe time it would take to perform this task manually.
The knock-on effect of this increased productivity is it Synbiosis reserves the right to amend or change
can ensure that making decisions about whether to specifications without prior notice. This Application
continue working with or discarding recombinants note supersedes all earlier versions.
becomes virtually instant. In some cases, this makes it possible for molecular biologists to undertake furtherprocessing of clones much sooner in the working day, thereby allowing results to be generated up to an entireday earlier.
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Matrix metalloproteinase Matrix metalloproteinases Collectively they are capable of degrading all kinds of molecules. They are known to be involved inin/activation. MMPs are also thought to play a major role on cell behaviors such as. They are distinguished from other endopeptidases by their s were described by Jermome Gross and Charles Lapiere (1962) who observed triple helix degrada

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