Oligo Donor Purification
Oligo Donor Purification - Related Videos
11:44
Synthesis and Mass Spectrometry Analysis of Oligo-peptoids
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Cited by 6 •
2018
A protocol is described for the manual synthesis of oligo-peptoids followed by sequence analysis by mass spectrometry.
08:43
OLIgo Mass Profiling (OLIMP) of Extracellular Polysaccharides
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Cited by 35 •
2010
A rapid way is described to gain insights into the structure of polysaccharides in an extracellular matrix. The method takes advantage of the specificity of glycosylhydrolases and the sensitivity of mass spectrometry allowing minute amounts of materials to be analyzed. This technique is adaptable to be used directly on tissue itself.
07:17
Single Port Donor Nephrectomy
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Cited by 5 •
2011
Single port laparoscopic surgery is changing the standard of care in surgical care like nothing since the laparoscopic technique was introduced 20 years ago. We present out technique of single port donor nephrectomy using the Gelpoint device. We have successfully performed this surgery in 100 patients.
12:13
Ex Situ Normothermic Machine Perfusion of Donor Livers
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Cited by 21 •
2015
Here we present a protocol describing oxygenated ex situ machine perfusion of donor liver grafts. This article contains a step by step protocol to procure and prepare the liver graft for machine perfusion, prepare the perfusion fluid, prime the perfusion machine and perform oxygenated normothermic machine perfusion of the liver graft.
08:16
Plasmid Purification
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2023
Plasmid purification is a technique used to isolate and purify plasmid DNA from genomic DNA, proteins, ribosomes, and the bacterial cell wall. A plasmid is a small, circular, double-stranded DNA that is used as a carrier of specific DNA molecules. When introduced into a host organism via transformation, a plasmid will be replicated, creating numerous copies of the DNA fragment under study. In this video, a step-by-step generalized procedure is described for how to perform plasmid purification.
FAQs
Purification improves donor quality by removing or reducing sequences that are not full length. Synthetic oligonucleotide production generates incomplete and chemically modified species alongside the intended sequence, so an unpurified preparation may contain molecules that do not provide the desired repair information. Enriching the full-length product supports more consistent use as a DNA repair template in genome-editing experiments.
Separation depends on measurable molecular properties rather than the intended sequence alone. High-performance liquid chromatography and polyacrylamide gel electrophoresis can resolve oligonucleotide species according to differences in size, charge, and hydrophobicity. These separations allow the desired full-length molecules to be enriched relative to incomplete or chemically modified products, improving the composition of the donor preparation.
High-performance liquid chromatography and polyacrylamide gel electrophoresis represent different purification options for the same quality goal: enriching intact donor molecules. The available information supports comparing them through their shared separation principle, rather than identifying one as universally superior. Both approaches can separate oligonucleotide species with differences in size, charge, or hydrophobicity.
Sequence fidelity and donor integrity matter because homology-directed repair uses the oligonucleotide as a template for a precise genetic change. When the preparation is enriched for the intended full-length sequence, it provides a more reliable repair substrate. In CRISPR-based editing, donor quality can influence editing accuracy, efficiency, and reproducibility, making purification an important part of experimental design.
A practical workflow begins with synthetic oligonucleotide production, followed by separation of the intended full-length product from incomplete and chemically modified species. Purification by HPLC or PAGE produces an enriched donor preparation, which can then serve as the DNA repair template in molecular biology or genome-editing work. The central outcome is improved donor quality for downstream experiments.
The approach is especially useful when experiments require precise genetic changes rather than a mixed or unreliable donor population. In CRISPR-based genome editing, enriched preparations support more dependable homology-directed repair substrates. The same quality improvement is relevant to variant modeling and functional studies, where sequence fidelity and reproducible donor performance help researchers interpret the effects of engineered changes.