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.

BIS Bulletin No 7, April 2020. Past epidemics had long-lasting effects on economies through illness and the loss of lives, while Covid-19 is marked by widespread containment measures and relatively lower fatalities among young people. The short-term costs of Covid-19 will probably dwarf those of past epidemics, due to the unprecedented and synchronised global sudden stop in economic activity induced by containment measures. The current estimated impact on global GDP growth for 2020 is around -4%, with substantial downside risks if containment policies are prolonged. Output losses are larger for major economies.

BIS Bulletin No 8, April 2020. Information on local labour markets and Google searches can be used to construct a measure of the vulnerability of employment in different regions of the United States to the Covid-19 shock. Regional exposure to Covid-19 varies significantly, ranging from a low of 2% to a high of 98% of total local employment. We test for the usefulness of the Covid-19 exposure measure by showing that areas with higher exposure report more Google search queries related to the pandemic and unemployment benefits.

The National Book Festival Presents series was created to provide a book festival experience to lovers of the event on a year-round basis. Because these programs can no longer be held at the Library, we are offering a virtual multipart series, with authors who have written books about widespread diseases and the worldwide response to them.

Our ongoing celebration of the Library of Congress National Book Festival continues with crime writer Patricia Cornwell discussing "Red Mist,” her 19th Kay Scarpetta novel, on the Fiction & Mystery stage at the 2012 Festival.

Our ongoing celebration of the Library of Congress National Book Festival continues with two-time Pulitzer Prize-winning novelist Colson Whitehead discussing "Zone One" on the Poetry & Prose stage at the 2012 Festival.

Rural educator Frank Cyr had the vision and pull to force the nation to standardize the color of the ubiquitous vehicle

Open for visitors, these houses model upcycling at its finest

A prototype deployed in San Francisco Bay imagines the underside of a floating building as an upside-down artificial reef

The graphic designer is receiving a Lifetime Achievement Award from Cooper Hewitt for her recognizable computer icons, typefaces and graphics

British product designer Lucy Hughes has invented a biodegradable plastic made from fish offcuts

From gas stations to public libraries, these celebrity architect-designed buildings are worth a road trip

The Wisconsin-born architect's buildings helped turn the city he once called an 'inglorious mantrap' into the center of the world

Press release about the Basel Committee publishing consultation paper on revisions to the credit valuation adjustment risk framework, 28 November 2019.

BIS Press Release - Easing trade tensions lift sentiment: BIS Quarterly Review, 8 December 2019

Press release: CPMI report sets out considerations for developers of wholesale digital tokens, 12 December 2019

Press release about the Basel Committee launching the consolidated Basel Framework, 16 December 2019.

Markets Committee calls for wider adoption of global code of conduct for foreign exchange markets (Press release, 30 January 2020)

No global real estate market despite higher price synchronisation and growing role of international investors, central banks find (Press release, 18 February 2020)

The Bank for International Settlements (BIS) today announced two key appointments to the BIS Innovation Hub, a new initiative designed to support central bank collaboration on new financial technology. (Press release, 19 February 2020)

Basel Committee Press release "Basel Committee meets to review vulnerabilities and emerging risks, advance supervisory initiatives and promote Basel III implementationl", 27 February 2020.

BIS Press Release - Shaping the future of payments: BIS Quarterly Review, 1 March 2020

BIS statement, 16 March 2020.

Press release about Basel Committee assesses members' implementation of the Net Stable Funding Ratio and large exposures framework, 19 March 2020

CPMI Press release "Jurisdictions move towards full implementation of standards for financial market infrastructures", 8 April 2020.

CPMI Press release "How fintech can promote financial inclusion - a new report on the opportunities and challenges", 14 April 2020

It works like a very fine "molecular knob" able to modulate the electrical activity of the neurons of our cerebral cortex, crucial to the functioning of our brain.

When we find something particularly beautiful or impressive, we literally get big eyes: Our pupils dilate. The pupil controls how much light enters the eye and falls on the retina.

With a discovery that could rewrite the immunology textbooks, an international group of scientists, including the teams of Bart Lambrecht, Martin Guilliams, Hamida Hammad, and Charlotte Scott (all from the VIB-UGent Center for Inflammation Research) identified a new type of antigen-presenting immune cell.

The purpose of suffering may be mysterious, but the search for meaning is obligatory.

The closures were necessary. Chasing picnickers and closing paint aisles is petty overreach.

The last time Charlene Chiddenton saw her father in person was on March 7 — visitor restrictions at long-term care homes during the pandemic had kept them apart — and less than two months later, she was saying goodbye to her father by video call.

When the province eventually conducts its review on the response to COVID-19, the president of the Nova Scotia Nurses’ Union can think of one thing in particular that needs to be discussed.

The only daily TV news package to focus on Nova Scotians and their stories

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.

A Dalhousie University scientist and a NSCAD textile professor are teaming up to create a new fabric. It could be used to locally manufacture personal protective equipment like surgical masks. As the CBC's Colleen Jones reports, they have received a COVID-19 grant to study the idea.

A number of fishing zones in the Gulf of St. Lawrence have been closed after North Atlantic right whales were sighted earlier this week.

Nova Scotia students and teachers will not be returning to the classroom this year. At-home learning will continue until June 5, when the province's school year will end.

Businesses in Cape Breton are helping customers get creative with DIY take-home kits to make everything from mermaid-themed terrariums to bubble tea.

Some school travel groups in Cape Breton that had trips cancelled in March due to COVID-19 are still waiting to get their money back.

A woman who lost her sister to COVID-19 at the start of the month is thanking the staff at Northwood who took "excellent" care of her in her final days. Jean Harrigan, 90, is being remembered for her caring and lively nature.

As Nova Scotians gazed up at the Canadian Forces Snowbirds soaring through the clouds last weekend, Matt MacKenzie was looking down and feeling the weight of successive tragedies that had enveloped his home province. 

Over the 13 hours the mass shooting took place across Nova Scotia on April 18 and 19, the RCMP called in resources from New Brunswick to help. However, the two municipal police forces closest to Portapique were not asked to respond directly at any point, CBC News has learned.

The latest death was at the Northwood long-term care home in Halifax.