Gibson Isothermal Assembly is a great technique for combining long double-stranded DNA fragments together. You can view a DIY protocol here, which works fine.
However, you should also know that you can buy all the components in a ready-to-use mastermix format from New England Biolabs (NEB). Details here.
IDT’s gBlocks are a great new product for anyone using traditional approaches to cloning or synthetic biology. Like any new technology, they take some getting used to. So, if you are a bit of a stick-in-the-mud, or a “late adapter” read the following and try to get inspired.
Easy Gene Assembly: gBlocks™ Gene Fragments (this article explains what gBlocks are)
Isothermal Assembly (how gBlocks can be assembled into full genes)
gBlocks User Guide (opens PDF file – all you need to know).
I am pleased to inform you (well, if you are a researcher in Denmark at least) of Agilent’s Summer promotion on genomics products. The promotion is valid from now(28 June 2011) until August 31st 2011. Headlines below, with a bit more detailed information beneath each heading
Update 12 July: Agilent have a Summer Promotion webpage now live.
Purchase the New SureCycle PCR Instrument Together with the Mx3005P qPCR Instrument and get 30% off the Bundle
This is possibly the best offer of the whole promotion. The SureCycler is the hottest new instrument from Agilent, very cool (connect to it from a mobile device) with great specs. The Mx3005P is a well-proven and robust qPCR instrument that won’t let you down.
Purchase the Bioanalyzer with Agilent SureCycler 8800 or with >5000€ of Bioreagents and get 30% off the Total Bundle
Hmm. On second thoughts, perhaps this the the best of the Summer Promos. It really depends on what you are after. Of course, Bioreagents can mean a lot of things: PCR enzymes/kits, nucleic acid sample prep and qPCR reagents would be the most obvious. The combination of the Bioanalyzer with qPCR reagents sounds like a good combination to me, given that the Bioanalyzer is great for doing QC on RNA.
Get 50% off Absolutely RNA extraction kit when you purchase Bioanalyzer reagents
For those of you who are reguarly ordering Bioanalyzer kits (and that means lots of you), this is a great opportunity to try out one of Agilent’s best nucleic acid purification lines. The Absolutely RNA extraction kits come in a number of size options (mini-, micro- and nano-prep) and there are kits for miRNA and FFPE samples too. This is really a great excuse for you to try out these kits, especially if you haven’t had a lot of success with what you are currently using.
10,000€ discount on your genomics consumables order when you purchase an Agilent Bravo Automated Liquid Handling Platform
OK, so not everyone has the money available for an Agilent Bravo. However, if you are in the market for a small-footprint, very fast and accurate pipetting workstation, you really have to consider the Bravo. You will save many many days of work by having a Bravo in your lab, not to mention saving your pipetting thumb!
10,000€ (about DKK 75K) will get you a lot of genomics consumables.
Genespring and exon arrays: buy 5000€ of exon arrays and get 25% off your Genespring annual license, buy 10k€ of exon arrays and get 50% off the annual license.
Yes, it is that simple. Purchase Agilent’s high-quality exon arrays and get a big saving on the GeneSpring license (which you can then use to analyse all that lovely data and put into a biological context).
So that is the summary of Agilent’s Summer Promotion on genomics products. It looks pretty good to me and will mean that you come back from your Summer holiday with new gear to get into your research again after the break.
Agilent are running a special Easter promotion that gives 25% or more discount on “Bioreagents” if ordered before April 31st, 2011. This is a short post aimed at researchers who might be interested in trying these reagents out by taking advantage of the offer. I have highlighted some tried-and-tested products in ten key application areas.
1. Application: High Fidelity PCR
When you really cannot afford errors.
Go for: PfuUltra II Fusion HS DNA Polymerase.
Here’s why: PfuUltra II is the highest fidelity enzyme out there and is available in a hotstart format. Capable of targeting up to 19kb.
Go for: Herculase II
Here’s why: Herculase II has outstanding performance for GC-rich templates – even with targets that contain as high as 85% GC content. Also has short extension times (15s) and can work with targets up to 12kb.
2. Application: Routine PCR
When you are looking for an economical price.
Go for: Paq5000 DNA Polymerase
Here’s why: Paq5000 has better sensitivity than Taq and can amplify much longer target sequences (up to 6kb). Also available as a hotstart mastermix, very competitively priced.
3. Application: Real-Time PCR (qPCR)
When you are looking to improve your results or need to run assays in the fastest possible time.
Here’s why: Brilliant III uses a new and unique Taq mutant that is almost certain to give you higher sensitivity and specificity than your current reagent.
4. Application: Reverse Transcription
When you are looking for high cDNA yield across a range of temperatures with low input RNA.
Go for: AffinityScript
Here’s why: AffinityScript has great performance across all temperatures from 37-55 °C. Excellent for low input RNA.
5. Application: RNA Isolation/Purification
When you are looking for pure, high-quality RNA from your samples, even down to single cells.
Go for: Absolutely RNA Kits.
Here’s why: Get high yields of intact, full-length DNA-free RNA. Flexible sizing to fit your requirements: Mini-, micro- and nano-prep kits. The last one of these has a 10µl elution volume and can isolate RNA from a single cell.
6. Application: Cloning
When you are looking for easy, reliable and affordable topoisomerase-based cloning.
Go for: StrataClonePCR Cloning Kits
Here’s why: Contains PfuUltra II DNA Polymerase for high accuracy and an easy, reliable protocol.
7. Application: Transformation of Competent Cells
When you are cloning large or difficult-to-clone DNA.
Go for: XL10 Gold Ultra-competent Cells
Here’s why: XL10 Gold cells were created specifically for transformation of large DNA molecules at high efficiency. Get 20-30 fold higher transformation than general cloning hosts.
8. Application: Transfection
When you want the same performance as the market leading product, but with significant cost savings.
Go for: GeneJammer Transfection Reagents
Here’s why: high transfection efficiencies with minimal cytotoxicity and you will probably save money
9. Application: Protein Expression in Mammalian Cells
When you are looking for affordable, fast and easy topoisomerase-based PCR cloning and expression.
Here’s why: uses a rapid protocol, with higher-fidelity enzymes than commonly found elsewhere.
10. Application: Site-Directed Mutagenesis
When you need tools to help you analyze protein functions, but don’t want to waste time on the molecular biology.
Go for: QuikChange Lightning Kits
Here’s why: QuikChange Lightning provides an accelerated protocol in a kit that is capable of handing short and long templates, all while maintaining the high accuracy, reliability and efficiency.
Researchers: get 50% off your first order when you open a Stratagene Virtual Freezer account with Agilent. Most people will ask the question “what is a Virtual Freezer?” In short, it is like having a departmental freezer, containing various reagents you use regularly, e.g. qPCR reagents, competent cells, cloning kits etc. However, in this case, Agilent keeps your products in a warehouse and ships directly to you when you need them. Delivery is 24 hours with free shipping. You can learn a bit more here, or by watching the video below.
Of course, if you have any questions, don’t hesitate to contact me.