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Related Concept Videos

Reproductive Cloning01:27

Reproductive Cloning

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Reproductive cloning is the process of producing a genetically identical copy—a clone—of an entire organism. While clones can be produced by splitting an early embryo—similar to what happens naturally with identical twins—cloning of adult animals is usually done by a process called somatic cell nuclear transfer (SCNT).
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The first successfully cloned mammal was Dolly, a sheep, born on 5th July 1996 at Roslin Institute, Scotland. The cloned sheep was named after the American singer Dolly Parton. Dolly lived for seven years and died of respiratory complications, which is speculated to be due to the actual age of her DNA. Because the DNA in cloned cells belongs to an older individual,  the cloned individual’s life expectancy may be affected. Indeed, analysis of Dolly’s DNA revealed shorter...
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DC Generator01:19

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An alternator converts mechanical energy into electrical energy that varies sinusoidally, resulting in AC current. Meanwhile, a DC generator converts mechanical energy into electrical energy, which are DC pulses with the same polarity. The construction of a DC generator is similar to that of an alternator, except that the pair of slip rings is replaced by a single split ring, also called a commutator. The commutator functions like a periodic rotary switch; it changes the contacts with the...
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The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold.
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Bacterial generation time, the period required for a bacterial population to double during its exponential growth phase, serves as a critical measure of microbial growth dynamics under optimal conditions. This parameter varies significantly across bacterial species and can be influenced by factors such as temperature, pH, and the availability of nutrients. For example, Escherichia coli can achieve a generation time of approximately 20 minutes, while Mycobacterium tuberculosis exhibits a much...
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Electric Generator: Alternator01:25

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Electric generators induce an emf by rotating a coil in a magnetic field. A simple alternator is an AC generator that creates electrical energy that varies sinusoidally with time. A simple alternator consists of a conducting loop that is placed inside a uniform magnetic field. The loop is connected to split rings connected to the external circuit with the help of brushes.
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Updated: Feb 16, 2026

Standardized Modular Assembly of Polycistronic Operons with Modular Cloning (MoClo) using the In-Cloning toolkit
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Standardized Modular Assembly of Polycistronic Operons with Modular Cloning (MoClo) using the In-Cloning toolkit

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Using Multisite LR Cloning to Generate a Destination Clone.

John S Reece-Hoyes, Albertha J M Walhout

    Cold Spring Harbor Protocols
    |January 4, 2018
    PubMed
    Summary
    This summary is machine-generated.

    This protocol details a multisite Gateway cloning method to combine a promoter and an open reading frame (ORF) into one Destination vector. This recombination technique enables the creation of diverse clones for various research applications.

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    Area of Science:

    • Molecular Biology
    • Biotechnology
    • Synthetic Biology

    Background:

    • The Gateway recombinatorial cloning system offers a versatile method for DNA manipulation.
    • Multisite cloning allows for the simultaneous transfer of multiple DNA fragments into a single vector.
    • Efficient generation of diverse expression constructs is crucial for various biological studies.

    Purpose of the Study:

    • To describe a protocol for multisite Gateway cloning.
    • To demonstrate the simultaneous transfer of a promoter and an open reading frame (ORF) into a Destination vector.
    • To generate a Destination clone with a fused promoter-ORF-GFP construct.

    Main Methods:

    • Utilizing the Gateway recombinatorial cloning system with LR enzymes.
    • Performing a multisite cloning reaction to transfer inserts from two Entry clones into one Destination vector.
    • Employing the pDEST-MB14 vector for creating a specific Destination clone.

    Main Results:

    • Successfully generated a Destination clone by simultaneously transferring a promoter and an ORF.
    • The cloned ORF was fused in-frame with a carboxy-terminal green fluorescent protein (GFP) moiety.
    • The protocol provides a framework for generating a wide array of Destination clones.

    Conclusions:

    • Multisite Gateway cloning is an effective strategy for constructing complex expression vectors.
    • This method facilitates the rapid generation of diverse clones for research.
    • The described protocol serves as a valuable guide for other multisite cloning applications.