Ultrasonic ligation for rapid high-efficiency subcloning in plasmid vectors

Cooper, J. B.; Halverson, Dana L.; Coldren, Chris D.

doi:10.1093/nar/21.7.1681

Cited by 5 publications

(1 citation statement)

References 8 publications

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“…DNA fragments were recovered from agarose gels using GeneClean (Bio-101 Co., La Jolla, CA) and ligated to vectors at 16OC either overnight or for 15 min in an ultrasonic bath (also at 16T) using T4 DNA ligase (Cooper et al, 1993). Ligation products were transformed into E. coli XL-1Blue competent cells.…”

Section: Dna Manipulations and Sequence Analysismentioning

confidence: 99%

A new proline-rich early nodulin from Medicago truncatula is highly expressed in nodule meristematic cells.

et al. 1994

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We cloned and characterized MtPRP4, a new member of the repetitive proline-rich protein gene family in Medicago truncatula. The sequence of MtPRP4 predicts a 62-kD protein consisting of a 22-amino acid N-terminal signal peptide and a 527-amino acid repetitive proline-rich domain composed of three repetitive pentapeptide motifs arranged into two decapeptide repeats: PPVEKPPVHK and PPVEKPPVYK. MtPRP4 is the largest PRP described to date and contains repeated motifs that have not previously been found together in a single polypeptide. RNA gel blot experiments detected MtPRP4 transcripts in symbiotic root nodules, but not in roots, hypocotyls, or leaves. Accumulation of MtPRP4 transcript was an early response to Rhizobium inoculation and did not depend on nodule infection. In situ hybridization experiments demonstrated that MtPRP4 was expressed early in the development of the nodule meristem and that expression was highest in the meristematic cells of mature indeterminate nodules. These data support the proposition that an important early response of legume host roots to Rhizobium involves remodeling the host extracellular matrix and that proline-rich wall proteins play an important role in this architectural modification.

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Section: Dna Manipulations and Sequence Analysismentioning

confidence: 99%

A new proline-rich early nodulin from Medicago truncatula is highly expressed in nodule meristematic cells.

et al. 1994

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Cook Your Samples: The Application of Microwave Irradiation in Speeding Up Biological Processes

Chen

Liu

et al. 2018

Mol Biotechnol

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Classic and conventional procedures in molecular cloning are inherent compositions in modern molecular biological experiments and are frequently involved in daily laboratory activities. They take up the majority of the total time input in spite of the availability of well-designed specialized commercial kits. A similar situation is also in the field of biotechnology. Fortunately, microwave/ultrasonic irradiation has been found to be capable of speeding up these processes, such as proteolysis in sample preparation for proteomics research, and digestion, ligation, (de)phosphorylation of DNA with the corresponding enzymes, even the introduction of DNA samples to recipient cells, and biotransformation (e.g., the production of biodiesel). Microwave/ultrasonic irradiation, when used solely or in combination with other existing operations, makes it possible to finish these time-consuming processes in as short as 1 min with comparable or even improved efficiency, and there is no need of reagent upgradation. The adoption of irradiation is ideal because it eliminates any possible side effects of the chemicals used as performance enhancer(s) that will inevitably make the system more complicated at least. More notably, the needed irradiation in the laboratory can be generated by a common microwave oven or ultrasonic cleaner. Taken together, microwave/ultrasonic irradiation provides an accessible method to make the procedures mentioned above time- and cost- efficient. In this article, we reviewed the relevant literature and discussed the experiment and mechanism details.

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Hengen

1993

Trends in Biochemical Sciences

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Ultrasonic ligation for rapid high-efficiency subcloning in plasmid vectors

Abstract: Numerous strategies have been developed for increasing the efficiency of ligation reactions using T4 DNA ligase for the production of recombinant plasmids. These include the addition

Cited by 5 publications

References 8 publications

A new proline-rich early nodulin from Medicago truncatula is highly expressed in nodule meristematic cells.

A new proline-rich early nodulin from Medicago truncatula is highly expressed in nodule meristematic cells.

Cook Your Samples: The Application of Microwave Irradiation in Speeding Up Biological Processes

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