Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

PCR01:32

PCR

210.7K
Overview
210.7K
DNA Isolation01:24

DNA Isolation

39.8K
DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...
39.8K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

A mechanistic study on the repair of cadmium-induced male infertility using Yishen Tongluo formula based on network toxicology and experimental validation.

Frontiers in endocrinology·2026
Same author

Corrigendum to "a healthy lifestyle is associated with lower risk of depression in type 2 diabetes, irrespective of genetic susceptibility: A UK biobank cohort study" [J. Affect. Disord. 405 (2026) 121657, doi:10.1016/j.jad.2026.121657].

Journal of affective disorders·2026
Same author

Multi-omics analysis of PFOA-induced sperm DNA damage in mice identifies candidate genes and mendelian randomization reveals their human genetic associations.

Reproductive biology·2026
Same author

The Detection of Radiation Effects in the Urine of Rhesus Macaques Using Raman Spectroscopy.

Radiation research·2026
Same author

LncRNA NORAD Promotes Spinal Cord Injury via miR-22-3p/PTEN Axis to Regulate Oxidative Stress and Inflammation in Neuronal Cells.

Global spine journal·2026
Same author

Engineering next-generation nanovaccines for maternal immunization and piglet protection against porcine epidemic diarrhea virus.

Journal of controlled release : official journal of the Controlled Release Society·2026
Same journal

Growth Model for Continuous Culture of a Hydrogen-Oxidizing Bacterium, Hydrogenophilus thermoluteolus Strain TH-1.

Biotechnology and bioengineering·2026
Same journal

Glycoengineered Recombinant Alpha1-Antitrypsin Results in Comparable In Vitro and In Vivo Activities to Human Plasma-Derived Protein.

Biotechnology and bioengineering·2026
Same journal

Minimizing Off-Target Effects of CRISPR-Cas9 With Optimized sgRNA: Evaluation of Efficiency and Specificity in the Tumor Protein 53 (TP53) Region.

Biotechnology and bioengineering·2026
Same journal

Metabolic Flux Analysis Reveals Cell Line-Specific Rewiring in CHO Cells Following TCA Cycle Intermediate Feeding for Bioprocess pH Control.

Biotechnology and bioengineering·2026
Same journal

Photohydrogenotrophic Cultivation of Purple Non-Sulfur Bacteria in an Open Bioreactor: Enhanced Selectivity Through Light Cycling and Ammonium Limitation.

Biotechnology and bioengineering·2026
Same journal

Translating Blue Light Stimulation From Batch to Perfusion: Process and Intracellular Metabolic Analysis.

Biotechnology and bioengineering·2026
See all related articles

Related Experiment Video

Updated: Aug 27, 2025

Use of In Vivo Assembly for High-efficiency Plasmid Construction
06:25

Use of In Vivo Assembly for High-efficiency Plasmid Construction

Published on: February 7, 2025

693

Lambda-PCR for precise DNA assembly and modification.

Imen Tanniche1,2, Amanda K Fisher1,3,4, Frank Gillam1,5

  • 1Department of Biological Systems Engineering, Virginia Tech, Blacksburg, Virginia, USA.

Biotechnology and Bioengineering
|September 23, 2022
PubMed
Summary
This summary is machine-generated.

Lambda-polymerase chain reaction (λ-PCR) is a new DNA assembly method. This technique offers high efficiency and stability for cloning projects, reducing costs and time compared to traditional methods.

Keywords:
DNA assemblyDNA modificationsPCRrecombinant plasmidsλ-exonuclease

More Related Videos

Improving Student Outcomes with an Adaptable Molecular Cloning Course-Based Undergraduate Research Experience
10:17

Improving Student Outcomes with an Adaptable Molecular Cloning Course-Based Undergraduate Research Experience

Published on: November 15, 2024

1.1K
Generation of Plasmid Vectors Expressing FLAG-tagged Proteins Under the Regulation of Human Elongation Factor-1α Promoter Using Gibson Assembly
10:18

Generation of Plasmid Vectors Expressing FLAG-tagged Proteins Under the Regulation of Human Elongation Factor-1α Promoter Using Gibson Assembly

Published on: February 9, 2015

37.2K

Related Experiment Videos

Last Updated: Aug 27, 2025

Use of In Vivo Assembly for High-efficiency Plasmid Construction
06:25

Use of In Vivo Assembly for High-efficiency Plasmid Construction

Published on: February 7, 2025

693
Improving Student Outcomes with an Adaptable Molecular Cloning Course-Based Undergraduate Research Experience
10:17

Improving Student Outcomes with an Adaptable Molecular Cloning Course-Based Undergraduate Research Experience

Published on: November 15, 2024

1.1K
Generation of Plasmid Vectors Expressing FLAG-tagged Proteins Under the Regulation of Human Elongation Factor-1α Promoter Using Gibson Assembly
10:18

Generation of Plasmid Vectors Expressing FLAG-tagged Proteins Under the Regulation of Human Elongation Factor-1α Promoter Using Gibson Assembly

Published on: February 9, 2015

37.2K

Area of Science:

  • Molecular Biology
  • Biotechnology
  • Synthetic Biology

Background:

  • Traditional DNA assembly methods can be costly and time-consuming.
  • Existing methods may struggle with large DNA fragments or complex assemblies.
  • The stability of single-stranded DNA (ssDNA) primers can be a limitation in PCR-based assembly.

Purpose of the Study:

  • To introduce and validate Lambda-polymerase chain reaction (λ-PCR), a novel open-source method for DNA assembly.
  • To demonstrate the advantages of using double-stranded DNA (dsDNA) intermediates for primer stability and cost reduction.
  • To showcase the versatility of λ-PCR across various cloning applications.

Main Methods:

  • Development of three λ-PCR variations: complete λ-exonuclease digestion, asymmetric PCR, and partial λ-exonuclease digestion.
  • Conversion of double-stranded DNA (dsDNA) products into single-stranded DNA (ssDNA) "megaprimers" for overlap extension.
  • Application of λ-PCR in four case studies, including simple cloning, multipart assemblies, and challenging gene cloning.

Main Results:

  • λ-PCR demonstrated high efficiency in assembling linear and circular DNA fragments.
  • The method proved advantageous for large DNA products, avoiding secondary structures and reducing costs.
  • Successful gene cloning was achieved in cases not possible with commercial kits.
  • Thermodynamic simulations were utilized to optimize λ-PCR assembly strategies.

Conclusions:

  • λ-PCR is an effective and versatile open-source tool for DNA assembly and cloning.
  • The method significantly reduces costs and time compared to conventional techniques and commercial kits.
  • λ-PCR offers enhanced stability and efficiency, particularly for complex or large DNA constructs.