Synthetic Biology:DNA synthesis order reviews

In an effort to collect information on the current state of DNA synthesis, please post recent experiences with DNA synthesis orders here. Details on the order are useful but not required. Any information is helpful. If you wish to post a review here, please request an account on OpenWetWare. Or alternatively, send your review to Reshma Shetty and I will post it here.

Information that might be useful includes

1. Your name and institution (optional, can post an anonymous review by emailing Reshma Shetty)
2. Order
   • Description
   • Approximate length
   • Difficulty (in your estimation, should this have been a hard order to fill or an easy one?)
3. DNA synthesis company
4. Experience
   • Cost (both estimated and final)
   • Turnaround time (both estimated and final)
   • Successfully synthesized, synthesized with mutations/errors or not synthesized?
5. Any other details that might be relevant

pSB4*5 BioBricks vector

1. Reshma Shetty, MIT
2. BioBricks vector components and vectors
   • 4 resistance markers (1kb each)
   • pSC101 origin (2kb)
   • BioBricks vector scaffold which contained both ccdB toxic protein and a pUC19 origin (2kb)
   • Assembly of 3 vectors (pSB4C5, pSB4T5 and pSB4K5; 4.5 kb each)
   • This order was very complicated. It included 2 replication origins and a toxic protein. Moreover, it involved both synthesis and assembly.
3. Codon Devices
4. Experience
   • Codon Devices had the cheapest quote for this order with prices as advertised on their website ($0.79/bp; $500 surcharge for clones over 1.5kb; and a surcharge for a custom E. coli strain). GENEART was only slightly more expensive. DNA 2.0 was significantly more expensive.
   • Delivery was estimated at 30-40 days. The 4 resistance markers took 6 weeks. The pSC101 replication origin took 8 weeks. The vector scaffold took 5 months.
   • The resistance markers and pSC101 replication origin were synthesized as ordered. The vector scaffold had some mutations (some due to their issues during synthesis and some possibly due to my design flaws). Codon Devices opted to not pursue the project further at this point and did not deliver on the 3 vectors. [Additional note: It turns out that the vector scaffold they delivered was actually two vector copies fused together in such a way that when cut by any single enzyme, it looked the right size. Only running the uncut vector on the gel revealed the unusually large fragment size.]
5. Codon Devices was very helpful as I set up and submitted this order which was critical since it was a complicated project. Over the course of the order, the technical team was responsive to questions and suggestions. Codon Devices' decision not to finish the order was frustrating (though they didn't charge for that part of the order). They didn't respond to an emailed complaint on this issue.

[As an added note, I went ahead and completed construction of these vectors on my own and they are now available via the Registry of Standard Biological Parts.]

IDT miniGenes

1. Austin Che, MIT
2. I've ordered a couple times from them, each less than 500nt ribozyme sequence
   • It is not being expressed so there should be no issues of cloning. It does have a lot of secondary structure in the RNA (and thus also in the DNA).
3. I looked at Geneart, DNA2.0, and IDT (miniGenes) and decided to go with IDT
4. Experience
   • Estimated cost = final cost = cost on their website ($95+$0.65/bp). Their cost is competitive with Geneart.
   • One order had extra $75 "complexity fee" due to stable hairpin secondary structure
   • No surprises at all and didn't need to go through online forms to "request quotes"
   • From order, to shipment, it was 6 business days! I had the plasmid in hand 10 days after ordering (even for order with added complexity fee due to hairpin.
5. Overall, very happy. For < 500nt sequences, IDT minigenes is excellent.