Hints and Tips for Working with RNA

How to Maintain an RNase-free Environment

An RNase free environment is essential when working with RNA samples. Obtaining full-length, high-quality RNA can be a real challenge in the lab.

The structure and lability of RNA, and the fact that enzymes that degrade RNA are ubiquitous, hardy, and nearly impossible to remove can make working with RNA difficult. Even trace amounts of RNase can degrade RNA, so it is essential to avoid inadvertently introducing RNase into RNA samples during or after the isolation procedure.

To help you ensure the best possible results when working with RNA, we’ve published some key hints and tips for working with RNA samples.

The article explores the main reasons for RNA degradation and discusses the various sources of RNases such as skin, dust, reagents and the samples themselves. It also includes suggestions on lab precautions you can take such as wearing gloves, using RNase inhibitors, and explains how high-quality reagents will help maintain an RNase-free environment.

Read Working with RNA: Hints & Tips →

Choosing the Right DNA Polymerase for PCR

Bioline scientists know what it takes to perform successful PCR. Their detailed understanding of enzymes and buffer chemistry drives our PCR technologies. In this white paper, Choosing the Right DNA Polymerase: A Guide to Successful PCR, they share their insights with you.

Find out about the four basic properties of DNA polymerases:

• Thermal Stability
• Extension Rate
• Fidelity
• Processivity

Learn how different configurations of these properties result in different classes of DNA polymerase: Thermostable, hot-start (HS), and high-fidelity, as well as polymerases for amplifying long amplicons.

The paper also covers the two crucial components of successful PCR – an optimized reaction buffer and a high-quality, thermostable DNA polymerase and explains how these attributes can help you choose the best enzyme for your research requirements.

Get the inside track on choosing the right DNA polymerase to ensure successful PCR by downloading your copy of the white paper today.

Download the White Paper →

2nd qPCR and Digital PCR Congress 2014, London

Bioline are pleased to be sponsoring and exhibiting at the 2nd qPCR and Digital PCR Congress in London, England on 20th and 21st October.

The meeting brings together key opinion leaders from industry and academia, and covers a range of important topics in real-time PCR, including:

• MIQE guidelines and standardization
• Quality control of real-time PCR assays
• Real-time PCR assay design, optimisation and validation
• Pathogen detection and quantification
• Sample preparation and quality control
• Single cell expression analysis
• Clinical and drug development applications
• miRNA, ncRNA, siRNA applications
• Bioinformatics and data analysis

Case studies in infectious diseases, vaccines, cancer, prenatal diagnosis, diagnostic and clinical applications, microbiology, food microbiology, plant and ecological genomics, as well as other novel applications will be presented.

A special session at the congress will be devoted to plant breeding, plant genomics and applied food and plant pathogen testing:

• Plant and ecological genomics
• Detection and identification of plant pathogens
• Gene expression analysis
• Real-time PCR in food research
• Food safety
• Genetically modified organism (GMO) quantification in food

Bioline maintains a significant real-time PCR research and development facility based in London, the heart of the MedCity biocluster for life science, which includes the new Francis Crick Institute.

See our Your Partner for Plant Research microsite to learn about some of our best tools to enhance your plant research experiments, and download our Plant Research Solutions Guide.

We will be showcasing our latest real-time PCR technological developments at the congress, including the new SensiFAST cDNA Synthesis Kit, a fast, easy-to-use method of generating the highest quality cDNA for highly accurate real-time PCR results.

Bioline will also be highlighting our industrial capabilities for custom manufacturing, including enzymes, buffer chemistries, dyes and internal controls for a wide range of applications from plant genotyping, agribiotech, pathogen testing and cancer research, to biomarkers and diagnostics research.

Our friendly team of real-time PCR specialists will be on-hand at our exhibition booth throughout the event and glad to answer your queries and discuss your research requirements.

For further details of speakers and topics, please see the Global Engage qPCR & Digital PCR Congress Agenda .

We look forward to seeing you at the 2014 congress.

Bioline’s Gem of an iGEM Offer is Back for iGEM2014!

Bioline is proud once again to offer its support to teams participating in the 2014 round of the annual iGEM competition. Teams supported by Bioline in previous years have won gold medals and advanced to the finals of the iGEM World Championships.

For a limited time only, all iGEM teams in the United Kingdom can take advantage of up to 40% off already highly-competitive Bioline list prices across our entire product range.

The Must-Have Bioline Toolbox for iGEM Researchers

Popular tools used by successful iGEM teams around the world include:

• Bioline Competent Cells and Cloning Reagents — Transform plasmids from the iGEM kit plates with Bioline’s extremely efficient alpha-select silver or gold efficiency cells.

We used bioline cells but didn’t follow the protocols because the cells were more competent so didn’t need to go through heat shock.— Boston iGEM 2012 Team

We also supply a handy range of antibiotics, SOC medium and a rather nifty quick stick ligase for both cohesive and blunt end cloning.

• MyTaq™ HS — Use MyTaq HS to amplify DNA from your minipreps in as little as 30 minutes, or amplify directly from your colonies! See the MyTaq colony PCR application note or this SOP written by the Danish iGEM2013 team.
• ISOLATE II Plasmid Mini Kit — Pure and very high DNA yields from your transformed colonies.
• VELOCITY DNA Polymerase — Perfect for all your high-fidelity applications.
• HyperLadder™ DNA Markers — Our hugely popular range of markers for DNA sizing, as used by Craig Venter’s research team!
• Real-time PCR Kits — Tried and trusted SensiFAST kits for gene expression analysis.
• Core Reagents — High quality buffers, agarose and other essentials.

All Bioline products are backed by our friendly and helpful UK based technical support staff, should you need it!

To request full details of our Gem of an iGEM Offer pricing plan, please contact your local Bioline UK account manager using our Rep Finder tool. Then, why not let your fellow iGem researchers know about this great offer using the sharing links below. Also feel free to Tweet or email us if you have any further questions

We look forward to hearing from you soon and we wish all iGEM2014 entrants every success in this year’s competition!

PS: You can also read about Bioline’s Synthetic Biology industry partnership with Liverpool Life Sciences UTC, the first school in the United Kingdom specialising in Science and Health Care for 14 to 19 year olds.

Terms and Conditions
This offer is valid on any product in our range for any iGEM team in the United Kingdom. No minimum or maximum spend to qualify for the discount. Expires on October 31, 2014. Not valid with any other promo codes or special offers. Please contact us for details of discounts available to iGEM teams in the United States, Germany, France, Singapore and Australia. Bioline products are also available through our carefully selected distributor partners.

Join Bioline at Analytica 2014 – Stand 516, Hall A3

Are you a scientist at the leading edge of biological discovery? Then join Bioline at the Analytica International Trade Fair in Munich, April 1-4, 2014.

Visit us on stand 516 in Hall A3 to explore our research solutions for gene expression and genotyping studies. Discover how our innovative products help you to achieve better and more accurate results.

This year we’ll be showcasing our new products SensiFAST™ cDNA Synthesis Kit, the unmatched MyTaq™ Blood PCR Kit, and the latest addition to our range, EPIK™ Kits for Epigenetics research and analysis.

Four live talks a day at Analytica 2014

We’ll be hosting live talks on the stand on all four days of the event, covering gene expression and cDNA synthesis, high-sensitivity qPCR, Bisulfite PCR, and all the latest innovations in Direct PCR.

Attend one of our talks most suited to your research, find out why Bioline is the perfect partner for your research, AND receive a free Bioline T-shirt!

10:30am — Superior Gene Expression
Improvements in cDNA Synthesis for qPCR
Better Reproducibility • Better Representation • Better Results

12:00pm — Sensitivity Matters
SensiFAST™: The Most Powerful qPCR Mix
Better Data • Better Results • Better Science
Exceed the Limit

2:00pm — Extreme PCR
Problems Solved with MyFi™ and MyTaq™
Bisulfite PCR with the new EPIK™ for Epigenetics kits
Bringing Reliability to Epigenetics

4:00pm — Latest Innovations in Direct PCR
New MyTaq™ Extract-PCR and MyTaq™ Blood PCR Kits
Learn more about the most sensitive genotyping kits around

We very much look forward to meeting with you at Analytica 2014!

Bioline Scholar – A CRISPR way of gene editing

An alternative way for RNAi libraries to modify the mammalian genome on a large-scale may lie in the use of genome editing technologies such as the use of targeted endonucleases. Such a technique includes Zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs) and clustered, regularly-interspaced, short palindromic repeats/CRISPR-associated endonuclease cas9 (CRISPR/Cas9). CRISPR is a Cas9 endonuclease–based method for sequence-specific genome modification. It can be used to facilitate efficient genome engineering in eukaryotic cells by simply specifying a 20-nt targeting sequence within its guide RNA.

These technologies have proven highly useful for gene editing genomes of various species and the speed at which CRISPR techniques have been exploited has been incredibly rapid.

In January 2013 researchers at MIT, the Broad Institute, and Rockerfeller University first reported the new technique based on the CRISPR/Cas9 system, for precisely altering the genomes of living cells through gene addition or deletion. Researchers say the technology could offer an easy-to-use and more cost-effective way to engineer organisms that produce biofuels; to design animal models to study human disease; or to develop new therapies, among other potential applications.

Bioline reagents have been used to augment the workflow in these exciting gene editing technologies to elucidate CRISPR systems, including our SensiFAST™ range of Real-Time PCR Kits, as well as our performance PCR polymerases.

SensiFAST™ No-ROX One-step Kit

In the first examples of genes that mediate the inhibition of a CRISPR/Cas system, researchers from the University of Toronto identified five distinct ‘anti-CRISPR’ genes in the genomes of bacteriophages infecting Pseudomonas aeruginosa.

Bondy-Denomy, J et al., (2012). Bacteriophage genes that inactivate the CRISPR/Cas bacterial immune system. Nature 493, 429-432.

MyTaq™ Red Mix

Researchers from Minnesota University reported precise, high-frequency editing of 15 genes in pig, goat, and cattle genomes using CRISPR/Cas and TALENs technology. RFLP analysis of colonies were analysed by PCR using MyTaq Red Mix.

Wenfang, T.W. et al., (2013). Efficient nonmeiotic allele introgression in livestock using custom endonucleases. PNAS 110(41), 16526-16531.

To learn more about performance PCR reagents from Bioline, just visit www.bioline.com

Bioline at BioTechnica 2013 – Delivering the latest in direct PCR amplification innovations

Bioline: The PCR Company, a global leader in molecular biology innovation, will be exhibiting (Hall, 9, Stand D26) and presenting (Hall 9, Stand F69) at Europe’s leading life science, biotechnology and laboratory show, BioTechnica 2013 (8-10 October, Hannover, Germany). Our Senior Global Product Manager, Dr. Steve Hawkins, will discuss the evolution of direct PCR amplification technologies as part of the prestigious Biotechnica Innovation Forum.

The recent increased focus on innovation in biotechnology and medical diagnostics within the world of life sciences has led to important advances in molecular detection and biomedicine. Bioline is proud to be at the forefront of some of the key developments in PCR technologies that are actively accelerating the pace and success of research for scientists around the world.

Analysis of DNA from blood: Technical challenges

Molecular screening of blood samples for blood-borne pathogens, blood cell irregularities and blood group genotyping is a routinely performed process, but can be fraught with stumbling blocks. Some of the key concerns when using PCR to analyse DNA isolated from blood cells include the potential for false-negatives, or poor results that arise from a lack of sensitivity induced by the presence of PCR inhibitors in blood. Common reaction inhibitors include hemoglobin, IgG, lactoferrin, proteases, anticoagulants and salts.

Fast PCR, direct from whole blood, without compromise

Our presentation at the Biotechnica 2013 Innovation Forum is titled ‘Fast PCR, Direct From Whole Blood, Without Compromise’. The talk will provide an overview of how technical challenges from direct PCR amplification of blood samples can be overcome using the very latest in direct PCR developments. We will outline an extremely rapid workflow that is specially engineered to overcome various PCR inhibitors typically present in whole blood samples that remove the need for pre-treatment or DNA isolation.

Ultra-fast PCR protocols specially developed to deliver significantly increased sensitivity and PCR success rates, without compromising quality and yield, will also be covered. We will also show how the speed and specificity of these techniques discussed make them the perfect solutions for performing high-throughput end-point multiplex PCR direct from whole blood.

Further details of our upcoming presentation at the Innovation Forum can be found on the Biotechnica 2013 web site.

Visit the brand new Bioline booth at Biotechnica 2013

Throughout the three days of Biotechnica 2013, we’ll be presenting all the latest innovations across our endpoint-PCR and real-time PCR product lines. If you’re attending, you’ll find the brand new Bioline stand in Hall 9, Stand D26 where you will also be able to enter our iPad prize draw. We invite you to experience the power of the latest in life science innovation with Bioline and look forward to seeing you there.

Bioline Sponsorship of the 19th Annual Conference of the Chinese Life Scientists Society in the UK

Bioline is proud to have served the Chinese research community in the UK for over two decades with high-quality reagents, and we’re proud to extend this support further by sponsoring the prestigious Chinese Life Scientists Society in the UK (CLSS-UK) 2013 Annual Conference.

The conference, to be held at the University of Oxford, 20-21 September, will bring together eminent Chinese and UK researchers from a variety of life science disciplines to share the latest industry advances, exchange ideas and to establish research collaborations.  Professor Andrew Hamilton, Vice-Chancellor of the University of Oxford, will give a welcome address to delegates, followed by Yang Shen, Minister Counsellor for Education at the Chinese Embassy in the UK.

Our performance product lines are available to researchers throughout China and Hong Kong via our trusted distribution partners and are also available throughout the Asia-Pacific region, including Japan, Korea, Taiwan, Thailand, Vietnam, Malaysia, Singapore and Indonesia, as well as in Australia and New Zealand via our Bioline Australia operation.

Bioline at the CLSS-UK 2013 Annnual Conference

Bioline’s product specialist, Dr Ray Chan, along with Bioline’s Account Manager for the University of Oxford, Manvir Tiwana, will be on hand at the conference to showcase our latest innovations for life science researchers and look forward to discussing the research requirements of attendees.

Additionally, to mark and celebrate the Chinese mid-autumn festival, Bioline will be running a special mooncake prize-draw – so don’t forget to visit the Bioline exhibition stand, avail yourself of some Bioline goodies, and enter the mooncake prize draw.  If you’re attending the conference, we’d love to hear from you, so feel free to drop us a line on Twitter.

Conference topics and areas of research to be covered this year will include:

• Biochemistry and Structural Biology
• Biotechnology
• Cancer Research
• Cardiovascular Bioscience
• Genomics and Proteomics
• Immunology / Virology / Inflammation / Infection
• Metabolic Diseases
• Molecular and Cell Biology
• Pharmacology
• Physiology
• Stem Cells and Neuroscience
• Translational Medicine and Clinical Science

Keynote speakers – two eminent scientists have been confirmed as the keynote speakers at this year’s conference:

• Duanqing Pei, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, China
• Terence Rabbitts, University of Oxford, UK

The following speakers have also been invited to the conference:

• Huiliang Li, University College London, UK
• Jilong Liu, University of Oxford, UK
• Pentao Liu, Wellcome Trust Sanger Institute, UK
• Paul Lu, University of Manchester, UK
• Shengjiang Tan, University of Cambridge, UK
• Zhidao Xia, Swansea University, UK
• Xiaoli Xiong, MRC National Institute for Medical Research, UK
• Luzheng Xue, University of Oxford, UK
• Chengcan Yao, University of Edinburgh, UK
• Bin Zhang, Icahn School of Medicine at Mount Sinai, USA

For full details, please see the official CLSS-UK event web site.

Hands-on Real-Time qPCR Workshop at Birkbeck, sponsored by Bioline

Quantifying nucleic acids by Real-Time qPCR
Hosted by
4^th-5^th September 2013, Birkbeck, University of London

Register now for a two-day, hands-on workshop sponsored by Bioline: The PCR Company that teaches you the core principles of qPCR and focuses on the practical steps required to design, set-up, validate and analyse Real-Time qPCR assays.

Teaching format: The workshop will focus on practical work by participants. Each session is prefaced by a brief lecture, with the timetable organised to encourage interaction, questions and discussion.

Suitable for: Researchers in academia, research institutes and the pharmaceutical and biotech industries who already have some familiarity with basic PCR principles.

Workshop venue: Birkbeck, University of London, in modern teaching labs in the heart of Bloomsbury, a short walk from Euston mainline station & several underground stations.

Workshop tutors: Dr David Sugden and Dr Patricia de Winter.

See full workshop details and register online.

Bioline is a leader in high-performance Real-Time qPCR reagents. To find out more about how Bioline SensiFAST™ Real-Time qPCR reagents can benefit your research, just visit bioline.com.

Synthetic biology innovation, building BioBricks with Bioline

The 6^th International Meeting on Synthetic Biology (hashtag: #SB6Conf), the world’s foremost synthetic biology (SynBio) meeting, is currently running in London, organised by the BioBricks Foundation, a non-profit organization based in Cambridge, Massachusetts.

BioBricks Foundation SB6.0 Co-Chairs include Professor Paul Freemont and Professor Richard Kitney of Imperial College, who will lead a new £10 million innovation and knowledge centre, to be called SynbiCITE, aimed at providing a bridge between academia and industry and announced at the conference by David Willetts, Minister for Universities and Science.

Imperial College is home to the UK Centre for Synthetic Biology and Innovation a £9m investment aimed at propelling the synthetic biology field forward and promoting SynBio start-ups. International collaboration and networking are important aspects of the meeting, along with poster presentations and ‘lightning talks’ from world leaders in the field of SynBio research.

David Willets said:

“Synthetic biology has huge potential for our economy and society in so many areas, from life sciences to agriculture. But to realise this potential we need to ensure researchers and business work together. This new Innovation and Knowledge Centre will help advance scientific knowledge and turn cutting edge research into commercial success.”

Professor Richard Kitney, co-academic of SynbiCITE added:

“Synthetic Biology could be the next ‘industrial revolution’ for the UK, where tiny devices manufactured from cells are used by us to improve many facets of our lives. From producing new, more sustainable fuels to developing devices that can monitor or improve our health, the applications in this field are limitless.”

The exciting and emerging field of Synthetic Biology research combines the disciplines of engineering and molecular biology to design and build novel, biologically-based parts, devices, and sensors, as well as the re-engineering of existing, natural biological organisms. Synthetic Biology has the potential to deliver important new applications, from detecting the early onset of disease and improving existing industrial processes, food production, green fuels, and developing therapies to fight harmful bacterial infections or cancers.

Much of the future success of synthetic biology is incumbent upon the development of standardized SynBio components that can be combined in predictable and repeatable ways. The precise approach used when fabricating a BioBrick component depends on the fabrication method (PCR or direct synthesis) as well as the type of part being constructed (a standard part or protein coding sequence).

SyntheticBiology.org maintains a nice introduction to Synthetic Biology as well as a useful how-to guide on the subject of constructing novel BioBrick parts for submission to the Registry of Standard Biological Parts.

Bioline makes it easy to harness the power of new generation enzymes to create parts for BioBricks using PCR. We manufacture and supply a range of high-performance PCR and molecular biology cloning tools, enabling researchers to drive their synthetic biology projects forward. Some of our most popular, most frequently used products for leading synthetic biologists presenting at SB6.0 (1, 2) include the High-Fidelity Velocity DNA Polymerase, MyTaq HS DNA Polymerase for Colony-PCR, Competent Cells, Quick-Stick Ligase for TA Cloning, ISOLATE II Plasmid Mini Kits, and our acclaimed range of SensiMix™ and SensiFAST™ Real-Time PCR kits.

If you’ve been attending the #SB6Conf in London this week, let us know if your poster cites any Bioline reagents and your synthetic biology research work and achievements could be showcased in a forthcoming SynBio article on the Bioline blog. And if you’re one of the IGEMers attending the conference, don’t forget to check out our Gem of an offer for iGEM Teams!

One final note regarding the future of synthetic biology and the synthetic biologists of the future, you can keep up to date with all the latest news from the iGEM SynBio research teams around the world by following the international iGEM Teams Twitter list maintained by us @ThePCRCompany.

Citations

1. Giuraniuc CV, MacPherson M, Saka Y, et al. (2013). Gateway Vectors for Efficient Artificial Gene Assembly In Vitro and Expression in Yeast Saccharomyces cerevisiae. PLoS ONE 8(5): e64419. doi:10.1371/journal.pone.0064419

2. Ali H, Ries MI, Nijland JG, Lankhorst PP, Hankemeier T, et al. (2013). A Branched Biosynthetic Pathway Is Involved in Production of Roquefortine and Related Compounds in Penicillium chrysogenum. PLoS ONE 8(6): e65328. doi:10.1371/journal.pone.0065328