Lemon 5-Min Cloning Package - Components - What’s included

Oct 22, 2026

Uploaded by asda1da

Lemon 5-Min Cloning Package provides components and protocols for rapid recombinant plasmid construction. The method mixes two DNA fragments with 15–18 bp identical terminal sequences and performs a room-temperature reaction of 3–5 minutes using Lemon Clonase 5x Premix. Included materials cover PCR master mix, DNA clean-up, transformation buffer, competent cell preparation, and glass beads for plating, enabling high transformation efficiency. Comparative results highlight strong cloning performance versus In-Fusion and Gibson methods, with a temperature stability test supporting maintained enzymatic activity.

LanguageEnglish

CategoryHealthcare

FormatPDF

Pages42

Reading Time1 h 46 min

AccessFree to download

Lemon 5-Min Cloning Package: Components

Lemon 5-Min Cloning Package: Advantages

Lemon 5-Min Cloning: Principle

Lemon 5-Min Cloning: Cloning Success Rate

Lemon 5-Min Cloning: Action Mechanism

Lemon 5-Min Cloning: Application

Lemon 5-Min Cloning: Advantages

Lemon 5-Min Cloning: Cloning Efficiency Comparison to other methods

In-Fusion vs Lemon 5-min

Gibson assembly vs Lemon 5-min

Test results

Lemon 5-Min Cloning: Temperature Stability Test

Lemon 5-Min Cloning Package: Components • Lemon 5-Min Clonase 5x Premix Tube (Store at-20ºC): 50 μL (50 reactions) • KOD One PCR Master Mix (2x) Tube from TOYOBO company (Store at-20ºC) : 1 mL (100 reactions) • PCR Product Clean-Up Kit (Store at RT): 100 reactions : DNA binding solution, Silica particle solution, Wash buffer • Transformation Buffer (TB) (Store at 4ºC): For making 100 ultra-efficient competent cell E-tubes • Glass Beads (Store at RT) : For plating. ► Before beginning your Lemon Cloning experiments, please read this Manual (Protocols) carefully, as well as the articles on the Naver Blog (Troubleshooting & Cloning Guidelines↑) . [https://blog.naver.com/vinejung](https://blog.naver.com/vinejung) ► For any Trouble-Shooting inquiry, please send an email to [vinejung@gmail.com](vinejung@gmail.com) [or](or genesandgrove@gmail.com)[ genesandgrove@gmail.com](or genesandgrove@gmail.com) Lemon 5-Min Cloning Package: Advantages 1. The Lemon 5-Min Cloning Method is fast (5-minute room temperature reaction), easy to use (one-step), and costeffective (1/5-1/10), requiring no temperature control equipment. It also enables the assembly of multiple DNA fragments, site-directed mutagenesis, DNA insertion/deletion, and simultaneous multiple cloning. 2. The KOD One PCR Master Mix included in the pack enables fast PCR reactions (5 seconds per 1 kb), highfidelity DNA amplification with a very low mutation rate, and supports the production of long PCR products (>10 kb) . Use this KOD polymerase only for high-quality DNA preparation with very low mutation rates for cloning purposes, not for colony PCR or genotyping. 3. The simple PCR clean-up kit in the pack yields highly concentrated DNA, which is essential for successful cloning. 4. The protocol for the ULTRA-efficient competent cells provided in the pack ensures very high transformation efficiency. 5. The Glass Beads included in the pack facilitate the easy spreading of E. coli cells on LB plates. Lemon 5-Min Cloning: Principle The New Gene Cloning Technology by Genes and Grove Lab The fastest gene cloning technology. By mixing two DNA fragments with 15-18 bp identical terminal sequences (at least 15 bp: 15-30 bp works well) and reacting them with Lemon Clonase 5x Premix at room temperature (20-30°C) for 3-5 minutes , a highly efficient recombinant DNA plasmid can be easily made. Lemon 5-Min Cloning: Cloning Success Rate By incubating the Lemon Clonase with DNA A and B (50 ng each) for only 5 minutes and transforming into C. cells, we obtained the following colony formation results even when only approximately one-sixth of the transformation culture was plated. This demonstrates a cloning success rate close to 100% . Lemon 5-Min Cloning: Action Mechanism Lemon 5-Min Cloning: Application Lemon 5-Min Cloning: Advantages The comparison of Lemon 5-min Cloning method with other cloning methods The Lemon 5-min cloning method is the fastest gene cloning technology. By mixing DNA fragments with identical terminal sequences (15-21 bp) and reacting them with cloning enzymes at room temperature for 3-5 minutes, a highly efficient recombinant DNA plasmid can be easily made. Moreover, multiple DNA assembly, sitedirected mutagenesis, DNA insertion/deletion, and high-throughput cloning can be efficiently done with this new method. Lemon 5-Min Cloning: Cloning Efficiency Comparison to other methods A. Comparison between the commercial In-Fusion cloning kit and Lemon 5-min cloning kit. The Lemon 5-min cloning kit demonstrates higher cloning efficiency compared to the In-Fusion method, requiring only a 5-minute enzyme reaction without the use of a temperature control device. A higher number of E. coli colonies indicates a highly efficient Lemon Cloning reaction. B. Comparison between the commercial Gibson assembly cloning kit and Lemon 5-min cloning kit. While the cloning efficiency is similar, the Lemon 5-min cloning kit has advantages in

FAQs

What DNA design is required for Lemon 5-Min Cloning?

Two DNA fragments must share identical terminal sequences of about 15–18 bp (at least 15 bp; 15–30 bp works well).

How long and at what temperature is the cloning reaction performed?

The reaction is carried out at room temperature (20–30°C) for 3–5 minutes.

How does Lemon 5-min cloning compare with In-Fusion and Gibson methods?

The document reports higher efficiency versus In-Fusion with a 5-minute enzyme reaction and similar efficiency versus Gibson, with advantages in speed and convenience.

Oct 22, 2026

Uploaded by asda1da