Ations for which there are no base analogs available. For instance, labeling with Cystamine hydrochloride can yield an internal thiol modifier to be used for crosslinking. Shown in Figure 2 is an example of a manual conjugation of Cystamine with an end-labeled NHS-dT. Upon reduction with DTT, a single peak was observed, indicating quantitative labeling.
Figure 1: The result of the On-Synthesizer labeling of the oligo 5′-6TT T6T TT6 TT-3′ where 6 = NHS-CarboxydT. Dansyl Cadaverine (4 mg/mL in 9:1 ACN/DMSO with 1% diisopropylethylamine) was pulled from port 6 and coupled for 5 minutes. The detailed procedure is described in the text.
These reaction schemes are detailed in Scheme 1 on Page 15. In addition, the reaction with propargylamine is shown to demonstrate the potential application to ‘click’ chemistry, as detailed on Page 9.162359-56-0 Biological Activity Conclusion This technique opens up the possibility of automating the synthesis of DNA probes labeled even with dyes that usually require post-synthesis labeling. A good example would be the popular AlexaTM dyes which are not compatible with regular oligonucleotide synthesis. However, these dyes are all available as either the cadaverine or hydrazine derivative (http://probes. invitrogen/handbook/tables/0728.html) and one could easily envisage automated high-throughput production of probes based on these dyes. In another example, labeling with propargylamine would offer an internal alkyne for Cu(I)-catalyzed `click’ chemistry conjugation with an azide. The possibility even exists to develop procedures for conjugating oligos with peptides and PNA derivatives.
FIGURE 2: RP HPLC OF CYSTAMINE-LABELED OLIGO
1) Cystamine Labeled Oligonucleotide
2) Unconjugated CarboxyModified Control
Figure 2: 1) The chromatogram of NHS-Carboxy-dT-T5 labeled with Cystamine-2HCl after reduction with DTT.6283-24-5 Description Cystamine-2HCl (15 mg) was dissolved in 1 mL DMSO with 35 DIEA added and allowed to react for 15 minutes at room temperature. After cleavage and deprotection in ammonium hydroxide, the disulfide was reduced with DTT and the oligo analyzed by RP HPLC.PMID:30480943 The earlier eluting peaks are DTT and oxidized DTT respectively. 2) The control oligo with the NHS-Carboxy-dT hydrolyzed with 1 M NaOH 15 minutes at 65 C. Note, when labeling an oligo internally with cystamine, the oligo must be capped with acetic anhydride/Nmethylimidazole after the cystamine conjugation to prevent branching off the second amine.
PRESORTED STANDARD US POSTAGE PAID RESTON VA PERMIT NO 536
GLEN-PAKTM CARTRIDGES THE ULTIMATE PURIFICATION CARTRIDGES
Introduction The use of Oligonucleotide Purification Cartridges has been a popular option for purifying synthetic oligonucleotides based on the DMT-on procedure. These cartridges typically contain polymeric packing materials which are stable to dilute ammonium hydroxide or ammonium hydroxide/methylamine (AMA). The technique relies on the affinity of reverse phase packing materials for the 4,4′-dimethoxytrityl (DMT) group at the 5′ terminus of synthetic oligonucleotides. The advantages of these cartridges are clear: speed, inexpensive, no need to evaporate corrosive solutions, product elutes in a small volume of aqueous acetonitrile. However, there is also a major disadvantage of traditional cartridges in that the longest oligos that can be reliably purified by this method are approximately 50mers. In addition, the yield of purified product from traditional cartridges is normally 40-60% of theoretical and the crude oligos need to be loaded.MedChemExpress (MCE) offers a wide range of high-quality research chemicals and biochemicals (novel life-science reagents, reference compounds and natural compounds) for scientific use. We have professionally experienced and friendly staff to meet your needs. We are a competent and trustworthy partner for your research and scientific projects.Related websites: https://www.medchemexpress.com
