DNA Synthesis
The Biopolymer/Genomics Core Facility has provided synthetic DNA services for over 20 years. High-through put, coupled to a rapid turn around time, with competitive pricing continue to attract our investigators and keep us competitive with commercial sources. Drs. Ambulos and Sadzewicz each have more than 15 years of experience in synthetic chemistry, including manual synthesis.
The facility has extensive experience with unusual chemistries including various antisense strategies (mixed or fully phosphorothioate, methyl phosphonate or H-phosphonate linkages), RNA synthesis and SiRNA synthesis. We have also gained extensive experience in the incorporation of modified bases (deoxy-uridine, 2-aminopurine, 5-methyl cytosine as well as custom synthetic phosphoramidites), fluorescent moieties (fluoresce in dyes, cyanin dyes, rhodamine dyes, as well asdual-labeled FRET probes), chemo-reactive modifiers (thiol and amino modifiers) as well as conjugation of oligonucleotides to synthetic peptides.
A large selection of modified bases, labels, and modifiers are routinely stocked in the facility. A partial listing of available modified bases, chemo-reactive modifiers and labels can be found at the link below. The service is supported by two Applied Biosystems instruments, a high-throughput 48-column model 3900 and a 4-column work-horse model 394 which is used predominantly for our modified bases and unusual or specialty chemistries.
Synthesis scales offered, range from 40nMole to 10mMole, although custom, larger scales have been accomplished using novel methods developed in our facility using our flexible peptide synthesis instrumentation. Turn around time for standard synthetic oligonucleotides is 24 hours, although sequences submitted in the morning are often processed and ready for delivery by that afternoon. Specialty oligos typically require an additional 24 to 48 hours. Purification capabilities include desalting or trityl-mediatedpurification by reversed phase HPLC. Rigorous quality control of synthetic oligos is accomplished by MALDI-TOF mass spectrometry.
| Scale | Cost per Base | Average Yield |
| 40nMole DNA | $0.40/base | 100-300μg |
| 0.2μMole DNA | $0.75/base | 600-1000μg |
| 1.0μMole DNA | $1.50/base | 1.5-2.0mg |
| 10μMole DNA | $10.00/base | 15-20mg |
HPLC purification is an available option. We use reversed phase HPLC methodology that takes advantage of the trityl protecting group used during synthesis to protect the 5' end. If HPLC purification is desired, we must know at the time we synthesize the oligonucleotide so that we can leave the final trityl group on, otherwise it is removed. HPLC purification costs are $50.00 per oligonucleotide synthesized at the 40nMole, 0.2μMole, or 1.0μMole scale. HPLC purification of oligonucleotides synthesized at the 10μMole scale is $100.00 per oligonucleotide. If you are performing trityl-based purification in your lab, please let us know at the time of ordering that you need the final trityl group left on the 5' end.
The Biopolymer/Genomics Core has a number of common primers that are 'Ready Made'. These oligonucleotides are HPLC purified, and provided in 10μg aliquots at 1μg/μl. Available primers include:
- Universal Forward
- Universal Reverse
- T7
- T3
- SP6
- Oligo-dT(20mer)
- Random Hexamer
Modified bases, labels, and chemical modifiers:
Refer to Glen Research and Biosearch technologies. What ever they have available in their catalog, we can incorporate! Follow the link below for a price list for many common modifiers and labels. If you don't see something listed, please contact us!
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