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Custom Gene Fragment Synthesis Services |
Invitrogen GeneArt Strings DNA Fragments are custom-made, uncloned, double-stranded linear DNA fragments assembled from synthetic oligonucleotides. They are an excellent choice for multi-fragment assembly with our GeneArt Gibson Assembly Cloning Kits. The low error rates of Strings DNA fragments mean less screening for correct, full-length sequence.
All Strings DNA Fragments can be optimized for protein expression at no extra cost and are also available as Strings DNA Libraries.
Order gene fragments with Strings DNA Synthesis Service
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- Configure projects, track order status, and get support all in one location
- Easily fix complex sequences with minimal changes
- Fully optimize sequence for improved protein expression
- Get free shipping for Strings DNA Fragments
Cloning fragments in high-throughput?
GeneArt HTP Strings DNA Fragments are an ideal choice for generating antibody variant expression libraries. Featuring a high quality, flexible format, they can be easily and efficiently integrated into your high-throughput cloning workflow.
Reasons to choose Strings DNA Fragments
Strings DNA Fragments are assembled from synthetic oligonucleotides using the same high-quality process developed for GeneArt Gene Synthesis. The synthesized DNA fragments are packaged in tubes and are delivered in a dried format, which can be readily resuspended, cloned, and screened for identification of the correct sequence. They do not contain adaptor sequences, eliminating the need to remove adaptors prior to use.
When using the GeneArt Dashboard to design DNA fragments, the proprietary Invitrogen GeneOptimizer algorithm optimizes sequences that can provide significant gains in downstream protein expression compared to wild-type sequences.
Benefits of Strings DNA Synthesis Service
- Affordable—GeneArt Strings DNA fragments are a cost-effective alternative to doing your own PCR or purchasing fully cloned genes
- Flexible—Sequence design flexibility with no adapters to worry about; use any downstream cloning method
- Fast—Improved turnaround times; at least 200 ng of GeneArt Strings DNA Fragments are produced in as little as 3–5 business days
- Optimized—Use our GeneArt GeneOptimizer software to improve downstream mRNA stability and increase protein expression by up to 15-fold over wild type
- Quality—Low error rates of <1:5,000 bp mean you screen fewer colonies to find the correct clone
Figure 1. Schematic of design and production of Strings DNA fragments.
New to GeneArt services? Explore GeneArt Dashboard quick guides
Production times for custom DNA fragments
Production time is the number of business days required to synthesize GeneArt Strings DNA products in our manufacturing facility.
| Fragment range (bp) | Production time (business days)* |
|---|---|
| 200–3,000 | 3–5 |
| DNA Libraries: IUPAC mixed bases | 10–15 |
* Delivery time is in addition to production time and depends on the destination of the shipment.
Tips for sequence design and screening of Strings DNA Fragments
Strings DNA Fragments can be cloned using any available cloning method, as long as the appropriate ends are included in the design. To identify a correct clone, screening of the clones is necessary. Find appropriate recommendations below.
For cloning compatibility, end sequences can be added to the Strings DNA fragment either before uploading the sequence into the GeneArt Services Dashboard or after import.
| Cloning method | Recommended sequence design |
|---|---|
| Restriction enzyme cloning | Add desired 5′ and 3′ restriction enzyme sequences, plus stuffer nucleotides on each end to ensure efficient enzyme binding |
| GeneArt Type IIs Assembly | Add desired 5′ and 3′ restriction enzyme sequences, plus stuffer nucleotides on each end to ensure efficient enzyme binding |
| Invitrogen GeneArt Seamless Cloning and Assembly | Add 15 bp sequence overlap (or more, depending on the kit) to vector or adjacent insert |
| Invitrogen Gateway Cloning | Add attB sites to enable proper recombination into Invitrogen pDONR vector |
| Invitrogen Zero Blunt TOPO PCR Cloning | Strings Fragments are blunt ended, so 5–10 stuffer nucleotides are recommended to compensate for small terminal truncations |
| TA and Invitrogen TOPO TA Cloning | Strings Fragments are blunt-ended and need to be adenylated using a Taq polymerase in the presence of dATP to add end-terminal A-overhangs |
Strings DNA Fragments are ready for screening to identify the correct clone. Strings DNA Fragments are bulk sequenced as part of the quality control process to help ensure that your desired sequence is present in the fragment pool. To identify a correct clone >90% of the time, we recommend using the following screening guidelines.
| Length of Strings Fragment | Screening recommendation |
|---|---|
| Up to 1 kb | Sequence 2 to 4 full-length clones |
| 1–2 kb | Sequence 3 to 5 full-length clones |
| 2–3 kb | Sequence 4 to 8 full-length clones |
Cloning options for Strings DNA Fragments: Performance data
GeneArt Gibson Assembly cloning kits
One to three Strings DNA fragments of 1 kb were assembled using the GeneArt Gibson Assembly HiFi Cloning Kit in pcDNA 3.4 vector using Invitrogen TOP10 competent cells (Figure 2).
GeneArt Gibson Assembly HiFi kits help provide high cloning efficiency using single- to multiple-insert designs.
Figure 2. Gibson Assembly of String DNA Fragments.
Restriction enzyme cloning example
Eight Strings DNA Fragments up to 3,000 bp were cloned using 5’ AscI restriction enzymes and 3’ PacI restriction enzymes. After ligation and transformation, bacteria were plated and incubated overnight at 37°C. Eight (<1 kb) or 16 (>1 kb) colonies were picked, and colony PCR was performed to identify full-length clones; four to eight full-length clones were sequenced to determine the number of sequence-correct clones.
The percentage of clones with correct length as identified by colony PCR (cloning efficiency) and the percentage of sequence-correct clones (fidelity) based on the number of full-length clones (Figure 3).
Figure 3. Restriction cloning of Strings DNA Fragments.
Cloning with GeneArt Type IIs Assembly
Eight Strings DNA Fragments of different lengths up to 2,800 bp with suitable sequence ends (AarI recognition sequence and 6 bp stuffer nucleotides) were used for direct assembly with the GeneArt Type IIs Assembly Kit, Aar I. The resulting four genes were 5,056 bp, 5,061 bp, 5,267 bp, and 5,448 bp long. After ligation and transformation, bacteria were plated and incubated overnight at 37°C. Sixteen colonies were picked, and colony PCR was performed to identify full-length clones. For all four assembled genes, eight colonies were sequenced to determine the number of sequence-correct clones.
The percentage of clones with correct length as identified by colony PCR (cloning efficiency) and the percent of sequence-correct clones (fidelity) based on the number of full-length clones (Figure 4).
Figure 4. Type IIs cloning of Strings DNA Fragments.
GeneArt seamless cloning
Ten Strings DNA Fragments up to 3,000 bp were cloned using the GeneArt Seamless Cloning and Assembly Kit. After ligation and transformation, bacteria were plated and incubated overnight at 37°C. Eight (<1 kb) or 16 (>1 kb) colonies were picked, and colony PCR was performed to identify full-length clones; Four to eight full-length clones were sequenced to determine the number of sequence-correct clones.
The percentage of clones with correct length as identified by colony PCR (cloning efficiency) and the percent of sequence-correct clones (fidelity) based on the number of full-length clones.
Figure 5. Seamless cloning with Strings DNA fragments.
Cloning two String Fragments with GeneArt Seamless Cloning and Assembly Kit
While Strings DNA Fragments are offered in lengths up to 3,000 bp, GeneArt Seamless assembly technology can be used to directly assemble two Strings DNA Fragments up to 1 kb without pre-cloning the subfragments.
If you want to assemble more Strings DNA Fragments or longer Strings DNA Fragments using seamless assembly technology, pre-cloning of the subfragments is recommended to limit post-assembly screening efforts necessary for finding correct clones. Alternatively, you can use GeneArt Type IIs assembly technology (Figure 6).
Figure 6. Direct cloning multiple Strings DNA fragments
Here, twelve Strings DNA fragments of different lengths up to 1 kb with suitable sequence ends (15 bp homology to the vector or to the next fragment) were used for direct assembly with the GeneArt Seamless Cloning and Assembly Kit. The resulting six genes were 1,375 bp, 1,466 bp, 1,536 bp, 1,590 bp, 1,647 bp, and 1,853 bp long. After ligation and transformation, bacteria were plated and incubated overnight at 37°C. Sixteen colonies were picked, and colony PCR was performed to identify full-length clones. For all six assembled genes, six colonies were sequenced to determine the number of sequence-correct clones.
Multi fragment assembly with Gateway cloning
Five Strings DNA Fragments up to 1,000 bp with attB sites were cloned into pDONR 221. After ligation and transformation, bacteria were plated and incubated overnight at 37°C. Eight colonies were picked, and colony PCR was performed to identify full-length clones; Four to seven full-length clones were sequenced to determine the number of sequence-correct clones.
The percentage of clones with correct length as identified by colony PCR (cloning efficiency) and the percentage of sequence-correct clones (fidelity) based on the number of full-length clones (Figure 7).
Figure 7. Gateway cloning with Strings DNA Fragments.
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