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Cat. No.: ECDP-400 (for 400T)

Cat. No.: ECDP-2k (for 2000T)

Description

E.coli Colony Direct PCR Kit is a convenient solution for performing PCR amplification with colonies or colony dilutions of Escherichia coli. This kit contains a 2X concentration of PCR pre-mix with blue and orange dyes (positioned at approximately 4 kb and 50 bp on electrophoresis). It includes 2X Taq DNA Polymerase, 2X PCR Buffer, 2X dNTPs, and 2X Loading Buffer. All that's needed is to add E. coli colonies or colony dilutions, primers, and water for the PCR amplification. Once amplification is complete, samples can be directly loaded onto electrophoresis gels. This kit is primarily designed for colony PCR screening and identification of positive clones following plasmid transformation in Escherichia coli.

Features

• This kit exhibits excellent compatibility with Escherichia coli colonies. Through repeated optimization, the E.coli Colony Direct PCR Kit ensures efficient PCR amplification directly from picked colonies, with minimal interference from colony presence on the PCR reaction.
• The kit offers exceptional convenience. The E.coli Colony Direct PCR Mix (Green, 2X) provided in the kit contains all the essential components, enabling users to perform PCR amplification of E. coli colonies with only the addition of primers and water. This simplifies PCR procedures, expediting the process, reducing contamination risks associated with PCR setup, and enhancing reproducibility. Additionally, the E.coli Colony Direct PCR Mix (Green, 2X) contains loading buffer components, allowing direct gel loading after PCR amplification without the need for additional loading buffer.
• The kit demonstrates robust amplification performance. It is capable of amplifying fragments up to 8 kb in length, though it is generally more suitable for amplifying DNA fragments below 2 kb.
• Dual-color dyes are included in the kit, facilitating visualization of the electrophoresis process. The addition of blue and orange electrophoresis tracking dyes (resulting in an overall green appearance) allows migration positions within a 1% agarose gel to be approximately 4 kb and 50 bp, respectively. After PCR, samples can be directly subjected to electrophoresis analysis without requiring additional loading buffer. The blue and orange tracking dyes do not interfere with the observation and detection of corresponding DNA bands.
• The product boasts high stability. Testing indicates that the E.coli Colony Direct PCR Kit maintains its amplification efficiency even after 15 repeated freeze-thaw cycles.

Storage

Store at -20°C for a minimum shelf life of one year. Avoid repeated freeze-thaw cycles. For frequent use, an aliquot can be stored at 4°C and will remain effective for at least 3 days.

Precaution

• Due to the high sensitivity of PCR amplification, which can increase the target gene sequence by over 10 million times, please take caution to prevent contamination of trace amounts of DNA to be amplified. Consider setting up negative controls without template DNA to confirm the absence of DNA contamination.
• The error rate of Taq DNA polymerase in each cycle of the PCR process is approximately 2.2×10^-5. For cloning DNA fragments larger than 1 kb, it is recommended to use DNA polymerases with lower error rates, such as Pfu DNA polymerase. However, for standard PCR or RT-PCR for qualitative or quantitative analysis, Taq DNA polymerase is the optimal choice.
• Despite maintaining nearly the same PCR amplification efficiency after 15 freeze-thaw cycles, it is still advisable to avoid repeated freeze-thaw of this product as multiple cycles might lead to a decline in product performance.
• Before using this product, ensure complete thawing and gentle inversion to mix thoroughly, while avoiding the formation of bubbles.
• This product is intended for scientific research by professionals only and must not be used for clinical diagnosis or treatment. It is not for use in food or drugs and should not be stored in regular households.
• For your safety and health, please wear appropriate laboratory attire and disposable gloves while handling this product.

Only for research and not intended for treatment of humans or animals

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