Synthetic genes for Agrigenomics

Applications

• Expression improvement in heterologous systems (e.g. codon usage adaptation)

• Cloning applications (e.g. cDNA cloning, qPCR standards)

• Optimisation of constructs (e.g. construction of hybrid genes)

Gene synthesis product portfolio

Our synthetic genes can easily be ordered through our online ordering platform. Integrated into this platform is our gene synthesis analysis and gene optimization tool, GENEius.

GENEius is a highly powerful tool for analyzing sequences online. If the sequence is complex, we will apply specific workflow steps to ensure the successful synthesis of the gene.

For non-complex sequences, we can ensure fast and cost-efficient sequencing using our highly automated workflow.

Non-complex sequences have:

• Moderate GC content (35-75%)
• No extensive repeats (>25 bp) or critical hairpin structures
• No extensive homopolymer stretches (>18 bp)

If you require your genes even more quickly, you can choose the "express option" during the final step of the ordering platform.

This will place your genes in a priority queue, and your gene will be synthesized in just 4 days.

Specifications for Express Genes:

• Miniprep scale only
• Valid for standard genes with moderate GC content (35-75%), without extensive homopolymer stretches (> 18 bp) and without extensive repeats (> 25 bp) or hairpin structures

Turnaround times for express genes

• 4 days: Genes up to 1000 bp, with no minimum base-pair count
• 6 days: Genes between 1001 - 1500 bp
• 7 days: Genes between 1501 - 2000 bp
• 11 days: Genes between 2001 - 3000 bp
• 13 days: Genes between 3001 - 4000 bp

Combinatorial library

Choose Gene Evolution libraries for screening projects to improve the properties of your protein of interest. We offer the following services:

• Mutations in single positions
• Mutations in multiple positions
• Mutations within specific domains
• Random substitution

Applications & Background

Gene Fragments are especially useful in applications such as gene assembly, mutagenesis studies, and DNA enzyme assays. They are also an ideal choice for sequences that exhibit high levels of toxicity in host cells.

GeneStrands are priced considerably lower than synthesised gene constructs making them a convenient choice in the construction of libraries and assembly of synthons.

Specifications

• Sequence length between 20 - 2000 bp
• Delivered dry as a dsDNA fragment (min. 100 ng)
  
• Non-complex sequences only
  
• Sanger sequence report upon request

The average turnaround time for GeneStrands up to 1000 bp is 5 - 7 working days; for GeneStrands > 1000 bp 8 - 10 working days.

If sequence validation is choosen: Electronic DNA sequencing data files will be delivered via Email.

Express GeneStrands

Express GeneStrands are the fastest product on the market. If you order until 2 pm CET your GeneStrands will be shipped the next day.

Only available for GeneStrands up to 1000 bp.

The synthetic linear dsDNA will be delivered as a fragment. If you order your Express GeneStrands until 2 p.m. CET we will ship them the next working day. If we cannot keep the next day shipment, we deduct the Express Fee from your invoice.

48h delivery available in all European countries and regions where UPS is offering Express shipment. Please check for your area: https://wwwapps.ups.com/ctc/request?loc=en_DE

Quality Assurance

All GeneStrands are synthesised under the same meticulous conditions as our gene synthesis service. However, GeneStrands are delivered as uncloned products, and as such the presence of minute populations of truncated or mutated species is unavoidable.

All GeneStrands are assembled from high-quality oligonucleotides synthesised at our in-house DNA synthesis facility. Assembled fragments are then size confirmed via electrophoresis.

Sequence-verified GeneStrands undergo DNA sequencing at our in-house sequencing facility. Sequence validation is of the entire pool, so the results obtained is a representation of the predominant sequence within the pool. Sequence verification is only possible for fragments ≥150 bp.

Eurofins Genomics routinely tests the mutation rate and quality of its synthesised Gene Fragments, and we have observed over 85% reliability of obtaining a correct clone if 6 or more colonies are picked and sequenced.

Plasmid DNA preparation services are required in research laboratories, biotech and pharmaceutical companies for applications such as:

• Production of viral vectors
• Production of antibodies and recombinant proteins
• Production of DNA vaccine
• Cell and gene therapy studies
• Other preclinical studies

Eurofins Genomics offers a variety of plasmid preparation services to cover individual needs:

Turnaround times

TAT for plasmid prep depends on availability of the QC'ed plasmid. Production can only start after we have received your plasmid and have run a QC, usually one Sanger DNA sequencing read. After successful QC the TAT is ~ 10 days. For large batches please inquire for available production slots before ordering.

TAT for plasmid prep services ordered in combination with gene synthesis is ~ 10 days.

Below you can find a wide range of possible SDM approaches

• Point mutations - a small number of base exchanges
  at one target site within a gene
• Insertions - from a few bases up to long strechtes of additional bases
• Deletions - remove unwanted parts of your gene
  sequence
• Large Substitutions - a longer stretch with several mutations at one target site
• Multi-position exchanges - alterations of the gene throughout many positions or the complete
  gene sequence

For all our SDM Services we offer

• Guaranteed confidentiality
  All confidentiality-sensitive steps are handled at our DNA campus: DNA Oligonucleotide synthesis, Sanger Sequencing and the SDM itself. Your data won't leave our Campus
• 100% accuracy of your mutated region
  All mutated sequences will be verified with double-stranded DNA Sequencing and a restriction digest for 100% accuracy
• Long term expertise in SDM
  The team in our lab has been working with SDM for many years and the routine performance of SDMs adds additional know how every day.
• Specific amplification method
  Our SDM is based on specific amplification methods to minimise errors and improve the quality

Our spectrum of cloning techniques can be used for a multitude of applications

• Cloning of full length cDNA
  Amplification of specific cDNA by RT-PCR and cloning into a vector of your choice
• Cloning of PCR products
  Amplification of specific DNA fragments by PCR and cloning into a standard vector or into a vector of your choice
• TA cloning
  Cloning of your PCR products into a TA cloning vector
  We clone any size of PCR product and your choice of polymerase is unrestricted
• Subcloning of vector inserts
  Subcloning of DNA fragments into target vectors, e.g. for in vivo protein expression, or for construction of hybrid genes
• Cloning of synthetic genes
  De novo synthesis of DNA fragments or complete genes and cloning into a standard vector or a vector of your choice, e.g. expression vectors

GENEius is a powerful tool to analyse, optimise and alter your sequence.

GENEius is directly integrated on our gene synthesis ordering platform

To learn more about GENEius, please click here >>

With our gene synthesis service, we offer a wide range of plasmid preparation scales and different options for your cloning vector.

Subcloning

Choose from several options for your cloning vector:

• Eurofins Genomics standard vector - for free -
• Send us your personal vector

Your gene is normally cloned in a standard vector for free. This vector contains either ampicillin or kanamycin resistance.

Plasmid Preparation

• Mini prep scale: 1-10µg plasmid DNA - for free -
• Midi prep scale: 15-50µg plasmid DNA
• Maxi prep scale: 100-500µg plasmid DNA
• Mega prep scale: 500µg-2.5mg plasmid DNA
• Giga prep scale: up to 10 mg plasmid DNA

You receive a free DNA mini prep that is carried out automatically on our robots. If the amount of DNA is insufficient, you can choose one of the scales above.

If you order either the mega or giga prep's, an endotoxin-free preparation procedure is available. For maxi prep scale Animal-free production is available.

Additional services

Linearisation of your gene

Linearisation of your gene is offered in combination with Midi and Maxi prep scales.

Completion of the digest will be checked via Agarose gel analysis. Some undigested plasmid might still be present in the final reaction.
For linearised mini- and midiprep plasmids the heat-inactivated digest reaction will be shipped.
Extra purification via spun columns for Mini or Midi preps is possible on request.
If the enzyme cannot be heat-inactivated additional spun column purification is available on request.
For Maxiprep alcohol precipitation is available. Shipment of pellet or linearised plasmid in solution is possible.

The standard service is available with the following enzymes: EcoR1, Nhe1, Not1, SacI, SaII, ScaI, XbaI, XhoI. Other enzymes are possible, but Eurofins will have to check if those enzyme can be made available free of charge. In some cases where Eurofins has to buy the enzyme, the enzyme price will have to be charged additionally.

Expert consultation (upon request)

We recommend our expert advice if your sequence template shows a high level of difficulties such as complex genes that contain many hairpins, repeats or other critical structures.

Sequence optimisation (upon request)

Using our proprietary software GENEius, the supplied amino acid or DNA sequences are optimised and adapted, e.g. for codon usage to any other organism. GENEius prevents direct or inverted repeats and hairpins and facilitates the distribution of G‘s and C‘s. Even the incorporation of "good motifs" is easily achieved with the GENEius software. Try our GENEius lightand discover what our optimisation software can do for you.

Experimental design

After submission of your desired sequence, we start the experimental design step. This is achieved by dividing the gene in overlapping oligonucleotides for gene synthesis by using GENEius.

Oligonucleotide synthesis

After design, the oligonucleotide synthesis phase begins. Our oligonucleotide synthesis facility - with more than 25 years of experience - guarantees highest quality and fast turnaround times.

Gene synthesis

Our gene synthesis lab is closely linked to the oligonucleotide production. The lab is the perfect combination of robotics and human know how. Aside from automation, our project managers, ensure best practise project layouts and stay in close touch with you if any question arise.

Subcloning

Based on your individual needs, your gene can be subcloned into one of our standard vectors or into any other vector of choice.

The standard vector maps can be found here. We use "a" high copy cloning vector with either ampicillin or kanamycin resistance.

More details about different vector options can be found under Prep Options & Subcloning.

Quality control

Quality that you can count on: our in-house sequencing facility guarantees 100% accuracy of your synthetic gene by double-strand DNA sequencing.

Delivery & plasmid preparation

For plasmid DNA preparation, a non manual mini-prep is generally performed free of charge as a standard procedure.
For higher yields, just order one of our plasmid preps yielding up to 10mg of your plasmid.
Endotoxin-free preparation for mega and giga scales are also available. Read more about our options for plasmid preparation in Prep Options & Subcloning

Documentation

We provide you with a quality assurance report and a delivery note after your synthetic gene has been synthesised via Ecom, our online ordering system: The quality assurance report contains:

• Detailed project specifications
• Complete plasmid description
• Restriction map
• DNA sequence information

Strict confidentiality is ensured by every project. The high quality of our service is overseen by a professional quality management. We are accredited according to DIN EN ISO 17025 and certified according to DIN EN ISO 13285 and GLP.