KOD Long PCR Master Mix　Code No.KML-101│TOYOBO PRODUCTS LIST│Research Reagents │TOYOBO

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DESCRIPTION

  • KOD Long PCR Master Mix is a 2 × PCR master mix formulated with genetically modified KOD DNA polymerase (UKOD). The combination of UKOD (known for its high fidelity and amplification efficiency) with an elongation accelerator enables rapid PCR at an elongation rate of 10 sec/kb. It maintains exceptional accuracy even with targets exceeding 10 kb. Thus, the master mix is ideally suited for amplifying long DNA fragments for long-read sequencing. It has a high success rate and also supports amplification from crude samples.
    KOD Long PCR Master Mix contains two types of anti-KOD DNA polymerase antibodies that inhibit both polymerase and 3′→5′ exonuclease activities, thus enabling Hot Start PCR. This master mix generates blunt-end PCR products because of the 3′→5′ exonuclease (proof-reading) activity of KOD DNA polymerase.

Features

- Ideal for Long-Read Analysis: Efficient Amplification of Long DNA Targets
KOD Long PCR Master Mix enables the amplification of DNA fragments up to approximately 50 kb using human genomic DNA as template. It is optimized for long-read sequencing applications, allowing the rapid and accurate amplification of long targets.

 

- Fast PCR
Targets ranging 1–10 kb can be amplified with an extension time of 5 sec/kb. Even for targets exceeding 10 kb, high-speed PCR is achievable with an extension time of 10 sec/kb, significantly reducing overall reaction time. The cycling conditions can be flexibly set when various targets of different sizes are to be amplified.

 

- High Fidelity
KOD Long PCR Master Mix exhibits approximately 80-fold higher fidelity than the Taq DNA polymerase. This high accuracy is particularly advantageous for various applications, such as the preparation of long target amplicons for sequencing.

 

- Compatible with GC-rich and AT-rich Targets
Amplification is possible regardless of the target sequence composition. With low amplification bias and high speed, KOD Long PCR Master Mix is also ideal for long-read library preparation.

 

- Amplification from Crude Samples
KOD Long PCR Master Mix is effective for amplification from crude samples (e.g., biological samples). Various sample types or lysates can be directly used as templates.

 
 
KML-101 Fidelity

Details

APPLICATIONS

  • ・Long read sequencing applications

SOURCE

E. coli strain carrying the cloned UKOD DNA polymerase gene

STORAGE CONDITION

Store at -20°C

COMPONENTS

KOD Long PCR Master Mix includes the following components for 200 reactions, 50 μl total reaction volume.

2x KOD Long PCR Master Mix 1 mL x 5

Application Data

Example 1.Amplification of long DNA targets (～47.5 kb) using fast PCR

Long DNA targets were amplified using KOD Long PCR Master Mix with human genomic DNA as template under fast PCR conditions.
Successful amplification of all long DNA targets was confirmed using this product.

application 1

Example 2.Long-Read Analysis of PCR Products

The 47.5 kb PCR product, amplified as described above, was purified and used to prepare a long-read sequencing library following the recommended protocol of the Ligation Sequencing Kit DNA V14 (Code: SQK-LSK114; Oxford Nanopore Technologies). The resulting library was analyzed using a Flongle Flow Cell (R10.4.1) (Code: FLO-FLG114; Oxford Nanopore Technologies).
Reads covering the entire 47.5 kb length were obtained, confirming that the target sequence was successfully amplified.

application 2