Fractionation of chromosomes II. Use of hydrophobic affinity partition in aqueous two-phase system

Bandyopadhyay, Dilip; Пинаев, Г. П.

doi:10.1016/0014-4827(83)90410-x

Cited by 6 publications

(2 citation statements)

References 13 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The selective uptake of DNA by dextranrich droplets was explored for the separation of single-and double-stranded DNAs, high-and low-molecular-weight DNAs, genomic DNA from a cell lysate, and even chromosomes. [15][16][17][18][19] We also observed that genomic DNA and DNA hydrogels were concentrated in the dextran-rich droplets, whereas short ssDNA were distributed in both dextran-rich droplets and a PEG-rich continuous phase. [20][21][22] We expected small ssDNA and large DNA structures would have different partitions and be separable by W/W-droplet fractionation (Figures 1B and 1 C).…”

Section: Introductionmentioning

confidence: 63%

“…In particular, a W/W‐droplet dispersion composed of phase‐separated dextran‐rich droplets dispersed in a PEG‐rich continuous phase has been widely used for DNA purification because of their selective uptake or exclusion of DNA molecules with specific physicochemical properties. The selective uptake of DNA by dextran‐rich droplets was explored for the separation of single‐ and double‐stranded DNAs, high‐ and low‐molecular‐weight DNAs, genomic DNA from a cell lysate, and even chromosomes [15–19] . We also observed that genomic DNA and DNA hydrogels were concentrated in the dextran‐rich droplets, whereas short ssDNA were distributed in both dextran‐rich droplets and a PEG‐rich continuous phase [20–22] .…”

Section: Introductionmentioning

confidence: 93%

See 1 more Smart Citation

Water‐in‐Water Droplets Selectively Uptake Self‐Assembled DNA Nano/Microstructures: a Versatile Method for Purification in DNA Nanotechnology

Masukawa

Sato

et al. 2022

ChemBioChem

View full text Add to dashboard Cite

DNA is an excellent material for constructing self-assembled nano/microstructures. Owing to the widespread use of DNA as a building block in laboratories and industry, it is desirable to increase the efficiency of all steps involved in producing selfassembled DNA structures. One of the bottlenecks is the purification required to separate the excess components from the target structures. This paper describes a purification method based on the fractionation by water-in-water (W/W) droplets composed of phase-separated dextran-rich droplets in a polyethylene glycol (PEG)-rich continuous phase. The dextran-rich droplets facilitate the selective uptake of self-assembled DNA nano/microstructures and allow the separation of the target structure. This study investigates the ability to purify DNA origami, DNA hydrogels, and DNA microtubes. The W/W-droplet fractionation allows the purification of structures of a broad size spectrum without changes to the protocol. By quantifying the activity of deoxyribozyme-modified DNA origami after W/Wdroplet purification, this study demonstrates that this method sufficiently preserves the accessibility to the surface of a functional DNA nanostructure. It is considered that the W/Wdroplet fractionation could become one of the standard methods for the purification of self-assembled DNA nano/ microstructures for biomedical and nanotechnology applications owing to its low cost and simplicity.

show abstract

Section: Introductionmentioning

confidence: 63%

Section: Introductionmentioning

confidence: 93%

Water‐in‐Water Droplets Selectively Uptake Self‐Assembled DNA Nano/Microstructures: a Versatile Method for Purification in DNA Nanotechnology

Masukawa

Sato

et al. 2022

ChemBioChem

View full text Add to dashboard Cite

show abstract

Isolation and fractionation of CHO chromosomes in aqueous two phase systems using charged polymers and base specific macroligands

Klaar¹,

Kula²

1986

Mol Cell Biochem

View full text Add to dashboard Cite

Chromosomes were isolated in a preparative scale by synchronisation of CHO cells with a double Thymidine block followed by an arrest in the metaphase by addition of Colcemid. Under proper cultivation conditions a mitotic index of 77% total cells could be routinely achieved. Bulk chromosome preparations free of nuclei and other subcellular particles have been obtained by low speed centrifugation followed by a 60 transfer countercurrent distribution using aqueous two phase systems composed of polyethylenglycol and dextran. The partition of CHO chromosomes previously purified in aqueous two phase systems were studied further to develop a protocol for the separation and isolation of individual chromosomes. Partition experiments with chromosomes changing the electrostatic phase potential by addition of charged PEG-derivatives suggest the existence of relatively highly charged chromosome groups. Most promising results with regard to separation were obtained using two PEG-derivatives, which interact specifically with the bases in DNA. For this affinity partitioning a GC- and AT-specific macroligand were employed. Comparing CCD's using each of these ligands information on the GC and AT content of exposed DNA in the chromosomes groups could be derived, demonstrating that specific sequences of DNA are accessible at the surface of metaphase chromosomes.

show abstract

Separation of rat testis cells by a manually operated countercurrent distribution apparatus

Bandyopadhyay

1984

Experimental Cell Research

View full text Add to dashboard Cite

Fractionation of chromosomes II. Use of hydrophobic affinity partition in aqueous two-phase system

Cited by 6 publications

References 13 publications

Water‐in‐Water Droplets Selectively Uptake Self‐Assembled DNA Nano/Microstructures: a Versatile Method for Purification in DNA Nanotechnology

Water‐in‐Water Droplets Selectively Uptake Self‐Assembled DNA Nano/Microstructures: a Versatile Method for Purification in DNA Nanotechnology

Isolation and fractionation of CHO chromosomes in aqueous two phase systems using charged polymers and base specific macroligands

Separation of rat testis cells by a manually operated countercurrent distribution apparatus

Contact Info

Product

Resources

About