When Elizabeth May, parliamentary leader of the federal Green Party, proclaimed on Tuesday that "oil is dead," she was correct in a philosophical sense. But not in a practical, real world sense.

Successful modification of the bacterial artificial chromosome (BAC) after two-step BAC engineering is confirmed in two separate polymerase chain reactions (PCRs). The first reaction (5' co-integrate PCR) uses a forward 5' co-integrate primer (a sequence located upstream of the 5' end of the A-box) and a reverse 3' primer on the vector (175PA+50AT) or within the reporter sequence or mutated region as appropriate. The second reaction (3' co-integrate PCR) uses a forward 5' primer on the recA gene (RecA1300S) and a reverse 3' co-integrate primer (a sequence located downstream from the 3' end of the B-box). Those colonies shown to be positive in PCR analysis are further tested for sensitivity to UV light. After the resolution, colonies that have lost the excised recombination vector including sacB and recA genes become UV light sensitive.

Bacterial artificial chromosome (BAC) clones are rendered electrocompetent and transformed with the recombinant shuttle vector, pLD53SCAB/AB-box. Cointegrates are selected by growth on chloramphenicol and ampicillin to ensure recombination of the shuttle vector into the BAC.

Plasmid DNA is prepared from the recombinant shuttle vector pLD53.SCAB/A-B created by cloning of the A and B homology arms for two-step bacterial artificial chromosome (BAC) engineering. To confirm that the A-box and B-box arms have been successfully incorporated into pLD53.SCAB, the pattern of enzyme digestion of the modified plasmid is compared with that of the unmodified pLD53.SCAB. Once the shuttle vector is shown to carry the proper sequences, it is ready for transfer into the BAC host.

This protocol describes the preparation of the shuttle vector before its introduction into bacterial artificial chromosome (BAC) host cells for BAC two-step engineering. The homology arm sequences, prepared previously, are introduced by ligation into the digested shuttle vector DNA to provide sites for recombination within the BAC clone. Crude lysates of individual bacterial transformants serve as templates in polymerase chain reaction (PCR) analysis to confirm the presence of the homology arms in the recombinant shuttle vector.

The 700-bp A homology arm (A-box) and the 700-bp B homology arm (B-box) are amplified by polymerase chain reaction (PCR) using purified bacterial artificial chromosome (BAC) DNA as template for two-step BAC engineering. The resulting A-box PCR product contains an AscI site at its 5' end (the 5' primer incorporates an AscI site, and the 3' primer does not incorporate any restriction sites). The B-box PCR product contains an XmaI site at its 3' end (the 5' primer does not incorporate any restriction sites, and the 3' primer incorporates an XmaI site). The amplification products are then digested with the appropriate restriction endonucleases to render them suitable for cloning into the shuttle vector.

In two-step bacterial artificial chromosome (BAC) engineering, a single plasmid is introduced into the BAC-carrying cell lines. The shuttle vector pLD53.SCAB (or pLD53.SCAEB) carries the recA gene and the R6K origin, which requires the protein to replicate. PIR2 cells, expressing , are typically used for the amplification of the vector and maintain about 15 copies/cell of the donor vector, which is relatively stable in this host.

Tissue engineering has a unique set of tools and technologies for developing preventive strategies, diagnostics, and treatments that can play an important role during the ongoing COVID-19 pandemic.

Classified released a music video for his song, Good News, this week featuring a pack of kids hanging out with seniors at a continuing care home in Enfield, N.S. It was filmed before the COVID-19 outbreak in the province.

Grim projections, travel rules and feeling safe while shopping at your local market — here's what's going on today.

"Recovery is not for the faint-hearted," says recovering addict Jeremy Raven. And sometimes, something as simple as a kind word may be the hand up that someone who is struggling needs, he says.

With something like 380 websites being created every minute, building an e-commerce shop can be a daunting task. It's easy to look at all of the success stories out there and think, "How could I possibly be that lucky?" However, the opportunity for growth in e-commerce is not slowing down. E-commerce sales are projected to grow 85 percent from their 2019 totals by 2022.

Voters have elected a new mayor in Flint, Michigan, where recovery from a lead-contaminated water crisis remains a major issue.

Almost every movie reminds Trump of … something it’s not.

The formative moment for a new generation.

Contribution by Mr Fabio Panetta, Member of the Executive Board of the European Central Bank, published by Politico on 21 April 2020.

CBC Ottawa's latest roundup of key points during the coronavirus pandemic.

More than 400 people have recovered from COVID-19 in Waterloo region, new numbers released by Region of Waterloo Public Health Thursday morning showed.

Four-time All Pro tight end Rob Gronkowski has agreed to a reunion with Tom Brady. The trade, finalized Tuesday night, brings Gronkowski and a seventh-round pick to Tampa Bay in exchange for a fourth-round selection.

B.C.'s unemployment rate grew by over four points in the month of April due to COVID-19, rising to 11.5 per cent.

Provincial Health Officer Dr. Bonnie Henry will give her daily briefing at noon.

With the effects of the pandemic playing out differently across the province, it's unlikely that all areas will reopen on the same timeline.

An inmate at the Calgary Remand Centre has tested positive for COVID-19, marking the first time a case has been reported at an Alberta correctional facility.

Prime Minister Justin Trudeau said Sunday that NHL players would — at a minimum — need to follow quarantine protocols if they were to arrive in Canada while the border remains closed due to the pandemic.

The post Danielle Boyer Makes STEAM Fun with The Engineering Design Club appeared first on SOLIDWORKS Education Blog.

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Penn State’s PROmoting School-community-university Partnerships to Enhance Resilience (PROSPER) project is continuing to connect and engage Pennsylvania families and keep schools informed during the COVID-19 pandemic.

Repair site iFixit has shared x-ray photographs of the new Magic Keyboard for iPad Pro, and they reveal an accessory more complicated than it might appear from the outside.

OM uses a team car to transport teenagers with autism to Montenegro's only special needs school. But a better car is needed to continue.

Almost one million people in Johannesburg, South Africa’s commercial hub, are in need of food aid due to movement restrictions imposed to curb the coronavirus pandemic, according to its mayor.

The post Perform Abaqus High-End Simulations from Home with Structural Mechanics Engineer appeared first on The SOLIDWORKS Blog.

The post An Electrical Engineer Using Sheet Metal? appeared first on The SOLIDWORKS Blog.

Workers deliver shoes to schools, bring food to the poor and help construct a kitchen for a family in the indigenous community of Chimaltenango.

We need a political system that mirrors the best in us.

These times are testing leaders from the schoolhouse to the White House, from city halls to corporate suites.

It will take more care than the president is currently demonstrating to loosen restrictions but still protect the vulnerable.

OM team in Hong Kong breaks cross-cultural ground, learning how to reach out to the ethnic minority, largely ignored by the Chinese population.

Four Nigerian marine engineering students studying in Cebu, Philippines, receive practical training and spiritual encouragement through a Logos Hope engineer.

After having some difficulty with the campers and teen leaders, OMer Stephen explains the significance of the cross and following Jesus—many respond positively.

After a trip to repair war damaged churches in the east, construction workers share their experience in order to inspire others to go.

Nirmal Purja, mountaineer

ANDY MURRAY may need to have another operation as he continues his bid to return to top-level tennis.

THE limit for in-store card transactions by contactless-enabled cards is to increase soon.

"In order to fulfil our mandate, there are several needs we must invest in," says Stephan Bauer.

"Do we sometimes feel uneasy that our business-as-mission and vocation models are so dependent on the very people that we are trying to reach? Does this make us feel powerless and needy?" asks Seang Pin.

Are you frightened of your garden? Terrified of killing your plants and overwhelmed by the sheer choice of species?

NEATEBOX, the Scottish technology firm which specialises in improving accessibility for people with disabilities, has been accepted into an accelerator programme run by Royal Bank of Scotland.

It wasn’t that long ago that health experts were championing golf as a soothing, morale-boosting haven away from the ravaging rigours of the coronavirus.