Automated seamless DNA co-transformation cloning with direct expression vectors applying positive or negative insert selection

<p>Abstract</p> <p>Background</p> <p>Molecular DNA cloning is crucial to many experiments and with the trend to higher throughput of modern approaches automated techniques are urgently required. We have established an automated, fast and flexible low-cost expression cloning approach requiring only vector and insert amplification by PCR and co-transformation of the products.</p> <p>Results</p> <p>Our vectors apply positive selection for the insert or negative selection against empty vector molecules and drive strong expression of target proteins in <it>E.coli </it>cells. Variable tags are available both in N-terminal or C-terminal position. A newly developed β-lactamase (ΔW290) selection cassette contains a segment inside the β-lactamase open reading frame encoding a stretch of hydrophilic amino acids that result in a T7 promoter when back-translated. This position of the promoter permits positive selection and attenuated expression of fusion proteins with C-terminal tags. We have tested eight vectors by inserting six target sequences of variable length, provenience and function. The target proteins were cloned, expressed and detected using an automated Tecan Freedom Evo II liquid handling work station. Only two colonies had to be picked to score with 85% correct inserts while 80% of those were positive in expression tests.</p> <p>Conclusions</p> <p>Our results establish co-transformation and positive/negative selection cloning in conjunction with the provided vectors and selection cassettes as an automatable alternative to commercialized high-throughput cloning systems like Gateway^®or ligase-independent cloning (LIC) <b/>.</p

Case studies of automation in services

A full understanding of the technological complexity underlying robotics and automation is still lacking, most of all when focusing on the impacts on work in services. By means of a qualitative analysis based on over 50 interviews to HR managers, IT technicians, workers and trade union delegates, this work provides evidence on the main changes occurring at shopfloor level in selected Italian companies having adopted technological artefacts potentially affecting labour tasks by automating processes. The analysis of interviews complemented with visits to the companies and desk research on business documents highlights that so far labour displacement due to the adoption of automation technologies is not yet in place, while tasks and organizational reconfiguration appear more widespread. Major heterogeneity applies across plants due to the final product/service produced, the techno-organizational capabilities of the firm and the type of strategic orientation versus technological adoption. These elements also affect drivers and barriers to technological adoption. Overall, the analysis confirms the complexity in automating presumably low-value-added phases: human labour remains crucial in conducting activities that require flexibility, adaptability and reconfiguration of physical tasks. Further, human agency and worker representation, in particular the role of trade unions, are almost disregarded and not considered by the firms when deciding to introduce a new technology

Case Studies of Automation in Services. A workplace analysis of logistics, cleaning and health sectors in Italy

A full understanding of the technological complexity underlying robotics and automation is still lacking, most of all when focusing on the impacts on work in services. By means of a qualitative analysis relying on the administration of more than 50 interviews to HR managers, IT technicians, workers and trade union delegates, this work provides evidence on the main changes occurring at shopfloor level in selected Italian companies having adopted technological artefacts potentially affecting labour tasks by automating processes. The analysis of interviews complemented with visits to the companies and desk research on business documents highlights that so far labour displacement due to the adoption of automation technologies is not yet in place, while tasks and organizational reconfiguration appear more widespread. Major heterogeneity applies across plants due to the final product/service produced, the techno-organizational capabilities of the firm and the type of strategic orientation versus technological adoption. These elements also affect drivers and barriers to technological adoption. Overall, the analysis confirms the complexity in automating presumably low-valueadded phases: human labour remains crucial in conducting activities that require flexibility, adaptability and reconfiguration of physical tasks. Further, human agency and worker representation, in particular the role of trade unions, are almost disregarded and not considered by the firms when deciding to introduce a new technology

Exploring individual user differences in the 2D/3D interaction with medical image data

User-centered design is often performed without regard to individual user differences. In this paper, we report results of an empirical study aimed to evaluate whether computer experience and demographic user characteristics would have an effect on the way people interact with the visualized medical data in a 3D virtual environment using 2D and 3D input devices. We analyzed the interaction through performance data, questionnaires and observations. The results suggest that differences in gender, age and game experience have an effect on people’s behavior and task performance, as well as on subjective\ud user preferences

Analysis, Design, and Implementation of a training center for variable-speed drive assembly production : Case ABB Oy

In manufacturing constant developments in production, processes, and layouts are required to respond towards increased production volume, quality, and customer requirements while meeting production targets and objectives. The case company of this thesis is ABB Ltd Drives Manufacturing Unit, which specializes in variable-speed drive production. ABB has recognized the need for re-designing a new and effective training center that supports One-piece flow assembly production since the old model is based on a cell production method. The training center is used for the training and integration of the company's new and experienced assemblers. The aim of the research is to analyze the current training concept, design a new technical solution, and create a detailed implementation plan. Thus, the following research questions were developed: RQ1: How to develop and re-design a training center that supports the assembler for One-piece flow method production of variable-speed drives? RQ2: How to design and create the best possible layout and solution to guarantee safety, flexibility, ergonomics, clear flow, and the maximum utilization of space? RQ3: How to implement a training center that does not disrupt the main production lines and makes that way operations more efficient? To achieve the objectives, the waste, bottlenecks, and issues of the current design were first identified by observing the training process and organizing focus groups and workshops with the production line and logistics (customer), and with the project team. Work-time studies were also conducted to solve the flow, outputs, cycle time, and waste time of the current process. These data collection methods aided in identifying potential improvement opportunities for the new design. The layout design process was committed by utilizing Lean principles and the Systematic layout planning procedure. AutoCAD was used to create and map various layout structures, options, and alternatives. The design process required the tendering of two layout location options, which were solved using the quantitative multiple attribute decision-making method, Weighted decision matrix (WDM), with voting based on the scoring of various criteria and features. The result was a Flexible 6-phase U-model one-piece flow training center that allows assemblers to be trained in both one-piece flow and cell production methods. The new design's scope of work was delivered to the supplier, numerous negotiations were held to achieve the best final solution, and the new training center was ordered. In the end, a detailed implementation plan with an estimated schedule was created and a future action list was established. The new design fulfils the objectives and eliminates all issues, waste, and bottlenecks while also ensuring safety, ergonomics, flexibility, a clear flow, and a high-quality training process. With the new design, the efficiency, quality, and output of training and production operations will improve.Teollisuuden alalla tuotantojärjestelmiä, prosesseja ja layouteja on jatkuvasti kehitettävä sekä modifioitava reagoidakseen kasvaneisiin tuotantomääriin sekä laatu- ja asiakasvaatimuksiin ja saavuttaakseen asetetut tuotantotavoitteet ja päämäärät. Tämän opinnäytetyön toimeksiantaja on ABB Oy Drives Manufacturing -yksikkö, joka on erikoistunut taajuusmuuttajatuotantoon. Toimeksiantaja on tunnistanut tarpeen uuden ja tehokkaamman koulutuslinjan suunnitteluun One-piece flow malliseen taajuusmuuttajien kokoonpanotuotantoon, sillä vanha tuotantomalli perustuu solutuotantomenetelmään. Koulutuslinjaa käytetään niin uusien kuten jo talossa olevien vanhojen kokoonpanoasentajien koulutukseen ja integrointiin. Tutkimuksen tavoitteena on analysoida nykyinen koulutuskonsepti, suunnitella uusi tekninen ratkaisu ja laatia yksityiskohtainen implementointisuunnitelma. Tavoitteiden saavuttamista varten on kehitetty seuraavat kolme tutkimuskysymystä: RQ1: Kuinka kehittää ja suunnitella koulutuslinja, joka tukee asentajia One-piece flow malliseen kokoonpanotuotantoon? RQ2: Miten suunnitella ja luoda paras mahdollinen layout ja ratkaisu, joka takaa turvallisuuden, joustavuuden, ergonomian, selkeän virtauksen ja maksimaalisen tilankäytön? RQ3: Kuinka implementoida koulutuslinja, joka ei häiritse päätuotantolinjoja ja tehostaa siten operaatioiden tehokkuutta? Saavuttaakseen tavoitteet, nykyisen koulutuskonseptin aiheuttamat pullonkaulat, ongelmat ja hukka tunnistettiin ensin havainnoimalla koulutusprosessia ja järjestämällä haastatteluja sekä työpajoja tuotantolinjan ja logistiikan (asiakkaan) sekä projektiryhmän kanssa. Nykyisen prosessin virtauksen, ulostulon, tahti -ja hukka-ajan selvittämiseksi suoritettiin myös työaikatutkimuksia. Nämä tiedonkeruumenetelmät auttoivat kehitysmahdollisuuksien tunnistamisessa uutta ratkaisua varten. Layout suunnitteluprosessi toteutettiin Lean-periaatteita ja systemaattista layout suunnittelua käyttäen. AutoCAD layout suunnittelusovellusta käytettiin erilaisien asettelurakenteiden ja vaihtoehtojen luomiseen sekä kartoittamiseen. Suunnitteluprosessi edellytti kahden layout-sijaintivaihtoehdon kilpailuttamista. Lopputulos ratkaistiin äänestämällä kvantitatiivisen päätöksentekomatriisin (WDM) avulla, joka perustui eri kriteerien ja ominaisuuksien pisteytykseen. Tulokseksi saatiin joustava 6-vaiheinen U-mallinen One-piece flow koulutuslinja, jonka avulla asentajia voidaan kouluttaa sekä One-piece flow että solutuotantomallisesti. Uuden koulutuslinjan työn laajuus -dokumentti toimitettiin toimittajalle sekä lukuisia neuvotteluja käytiin parhaan loppuratkaisun saavuttamiseksi, jonka jälkeen uusi koulutuslinja tilattiin. Lopuksi koostettiin yksityiskohtainen implementointisuunnitelma arvioituineen aikatauluineen ja laadittiin toimenpidelista tulevaisuutta varten. Uusi ratkaisu täyttää asetetut tavoitteet ja eliminoi kaikki ongelmat, hukat ja pullonkaulat sekä takaa turvallisuuden, ergonomian, joustavuuden, selkeän virtauksen ja laadukkaan koulutusprosessin. Uuden ratkaisun myötä koulutuksen ja operaatioiden tehokkuus, laatu ja tuottavuus paranevat

The future of laboratory medicine - A 2014 perspective.

Predicting the future is a difficult task. Not surprisingly, there are many examples and assumptions that have proved to be wrong. This review surveys the many predictions, beginning in 1887, about the future of laboratory medicine and its sub-specialties such as clinical chemistry and molecular pathology. It provides a commentary on the accuracy of the predictions and offers opinions on emerging technologies, economic factors and social developments that may play a role in shaping the future of laboratory medicine

Sensor-based navigating mobile robots for people with disabilities

People with severe physical disabilities need help with everyday tasks, such as getting dressed, eating, brushing their teeth, scratching themselves, drinking, etc. They also need support to be able to work. They are usually helped by one or more persona