Extraction of Nucleic Acid Fragments from Gels

Kurien, Biji T.; Scofield, R. Hal

doi:10.1006/abio.2001.5526

Cited by 18 publications

(17 citation statements)

References 54 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…5). The yield for large DNA was lower than that for short one, which was in accordance with the results reported before [1,3]. The yield for l DNA was relatively low.…”

Section: Yield Of Recoverysupporting

confidence: 93%

“…Agarose gels can resolve DNA from 150 to 880 000 bp at agarose concentrations ranging from 0.1-2.5% [3,8]. For preparative work by an agarose gel, the maximal amount of DNA compatible with a sharp, clean band is about 100 ng.…”

Section: Yield Of Recoverymentioning

confidence: 99%

“…For preparative electrophoresis, recovery and purification of the interest DNA from gel slices is necessary and plays an essential role in further manipulations, such as PCR, DNA ligation, cloning, sequencing and hybridization. Consequently, a myriad of methods have been developed for this goal over the years [1][2][3]. However, as far as speed, simplicity, yield, purity and cost are concerned, none of them has been entirely satisfactory and universally accepted as the norm procedure.…”

Section: Introductionmentioning

confidence: 98%

“…The first method involves the physical extrusion of DNA from the gel slices by centrifugal force through devices such as pipet tips [4], microcentrifuge tubes [5], filters [6], concentrators [7] and spin columns [8]. While centrifugation-based techniques are generally simple and rapid, the problems for them have been the requirement for the use of lower concentration agarose, which limits the separation of short DNA fragments [3]. In addition, centrifugation may also cause coelution of contaminating agarose or buffer component that interferes with subsequent enzymatic manipulations of DNA.…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Rapid recovery of DNA from agarose gel slices by coupling electroelution with monolithic SPE

Yang

Zhou

et al. 2009

Electrophoresis

View full text Add to dashboard Cite

An amino silica monolithic column prepared by in situ polymerization of tetraethoxysilane and N-(beta-aminoethyl)-gamma-aminopropyltriethoxysilane was firstly applied to recover DNA from agarose gel slices by coupling electroelution with monolithic SPE. DNA was electroeluted from the agarose gel slices onto the amino silica monolithic column. The DNA adsorbed on this monolithic column was then recovered using sodium phosphate solution at pH 10. The whole recovery procedure could be completed within 10 min because the use of amino silica monolithic column accelerated the DNA capture and facilitated the DNA release. Electroelution conditions, such as buffer pH, buffer concentration and applied voltage, were online optimized. The average yield for herring sperm DNA, pBR 322 DNA and lambda DNA recovered from 1.0% w/v agarose gel slices were 55+/-4, 50+/-6 and 42+/-7% (n=3), respectively. The polymerase chain reaction performance of pGM plasmid recovered from agarose gel slices demonstrated that the method could provide high-quality DNA for downstream processes. The combination of electroelution with monolithic SPE allows a rapid, simple and efficient DNA recovery method. This technique is especially useful for applications that need to purify small starting amounts of DNA.

show abstract

“…5). The yield for large DNA was lower than that for short one, which was in accordance with the results reported before [1,3]. The yield for l DNA was relatively low.…”

Section: Yield Of Recoverysupporting

confidence: 93%

Section: Yield Of Recoverymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 98%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Rapid recovery of DNA from agarose gel slices by coupling electroelution with monolithic SPE

Yang

Zhou

et al. 2009

Electrophoresis

View full text Add to dashboard Cite

show abstract

“…Essentially the same elution methods as described for proteins in Section 3 can be applied directly to nucleic acids. Because several reviews are dedicated specifically to nucleic acid elution (e.g., [262,263]), only a few examples are selected to emphasize the versatility of preparative electrophoresis.…”

Section: Elution Of Nucleic Acidsmentioning

confidence: 99%

Preparative isolation of protein complexes and other bioparticles by elution from polyacrylamide gels

Seelert

Krause

2008

Electrophoresis

View full text Add to dashboard Cite

Due to its unmatched resolution, gel electrophoresis is an indispensable tool for the analysis of diverse biomolecules. By adaptation of the electrophoretic conditions, even fragile protein complexes as parts of intracellular networks migrate through the gel matrix under sustainment of their integrity. If the thickness of such native gels is significantly increased compared to the analytical version, also high sample loads can be processed. However, the cage-like network obstructs an in-depth analysis for deciphering structure and function of protein complexes and other species. Consequently, the biomolecules have to be removed from the gel matrix into solution. Several approaches summarized in this review tackle this problem. While passive elution relies on diffusion processes, electroelution employs an electric field to force biomolecules out of the gel. An alternative procedure requires a special electrophoresis setup, the continuous elution device. In this apparatus, molecules migrate in the electric field until they leave the gel and were collected in a buffer stream. Successful isolation of diverse protein complexes like photosystems, ATP-dependent enzymes or active respiratory supercomplexes and some other bioparticles demonstrates the versatility of preparative electrophoresis. After liberating particles out of the gel cage, numerous applications are feasible. They include elucidation of the individual components up to high resolution structures of protein complexes. Therefore, preparative electrophoresis can complement standard purification methods and is in some cases superior to them.

show abstract

Sandcastle Worm‐Inspired Blood‐Resistant Bone Graft Binder Using a Sticky Mussel Protein for Augmented In Vivo Bone Regeneration

Kim

Choi

Jun

et al. 2016

Adv Healthcare Materials

View full text Add to dashboard Cite

Xenogenic bone substitutes are commonly used during orthopedic reconstructive procedures to assist bone regeneration. However, huge amounts of blood accompanied with massive bone loss usually increase the difficulty of placing the xenograft into the bony defect. Additionally, the lack of an organic matrix leads to a decrease in the mechanical strength of the bone-grafted site. For effective bone grafting, this study aims at developing a mussel adhesion-employed bone graft binder with great blood-resistance and enhanced mechanical properties. The distinguishing water (or blood) resistance of the binder originates from sandcastle worm-inspired complex coacervation using negatively charged hyaluronic acid (HA) and a positively charged recombinant mussel adhesive protein (rMAP) containing tyrosine residues. The rMAP/HA coacervate stabilizes the agglomerated bone graft in the presence of blood. Moreover, the rMAP/HA composite binder enhances the mechanical and hemostatic properties of the bone graft agglomerate. These outstanding features improve the osteoconductivity of the agglomerate and subsequently promote in vivo bone regeneration. Thus, the blood-resistant coacervated mussel protein glue is a promising binding material for effective bone grafting and can be successfully expanded to general bone tissue engineering.

show abstract

Extraction of Nucleic Acid Fragments from Gels

Cited by 18 publications

References 54 publications

Rapid recovery of DNA from agarose gel slices by coupling electroelution with monolithic SPE

Rapid recovery of DNA from agarose gel slices by coupling electroelution with monolithic SPE

Preparative isolation of protein complexes and other bioparticles by elution from polyacrylamide gels

Sandcastle Worm‐Inspired Blood‐Resistant Bone Graft Binder Using a Sticky Mussel Protein for Augmented In Vivo Bone Regeneration

Contact Info

Product

Resources

About