DeCodi-Fi typically requires 30 seconds to extend DNA fragments under 1 Kilobase (kb). For larger fragments, add 15 seconds(sec) per additional kilobase. For example, to amplify a 23 kb fragment should be 30 sec plus 15 sec x 22kb which is 360 seconds (6 minutes).

DeCodi-Fi amplifies high-complexity DNA starting from 0.1 ng template and medium-complexity DNA starting from 5 pg. 

Recommended input amounts are 5–50 ng for genomic DNA and less than 1 ng for templates smaller than 50 kb. To obtain amplification at 25 cycles, load more than 10⁴ copies of the template, but do not exceed 100 ng per reaction (e.g. 35 ng of human genomic DNA corresponds to 10⁴ copies).

Store the box at -20°C immediately upon receipt to maintain optimal kit performance and avoid repeated freeze-thaw cycles. 

DeCodi-Fi is designed by Kura Biotech to be transported at temperatures between 2°C and 8°C without performance loss. Tests have confirmed the components remain stable under these conditions for at least 10 days during transport.

The polymerase is inactivated by an antibody until the initial cycle of thermal denaturation. This prevents non-specific priming during setup and initiation, reducing unwanted amplification products and improving overall reaction efficiency.

DeCodi-Fi polymerase has higher amplification efficiency than commonly used polymerases, even for longer amplicons. We recommend starting with 25 cycles for most PCR with inputs >10⁴ copies of template.  Always keep cycling under 30, particularly for amplicons >10kb. Shorter amplicons can tolerate overcycling better. 

For amplifying Illumina sequencing libraries <600bp we recommend 12-14 cycles when starting from 1ng of template for reaching peak yield. 

The number of cycles can range from 10 to 30, with 25 cycles as a good starting point. Optimization may be needed based on your specific template, primers, and application. Fewer cycles reduce the probability of errors and help minimize nonspecific products, smearing, and the accumulation of high molecular weight artifacts in agarose gels.

Yes, we offer OEM and custom solutions that allow you to utilize our extensive expertise in enzyme optimization, buffer refinement and manufacturing. 

We recommend adding two phosphorothioate bonds at the 3' end of primers for avoiding 3' exonuclease degradation, given by the strong proofreading activity of DeCodi-Fi. This is particularly important for shorter primers or when amplifying NGS libraries. 3' primer degradation could result in amplification of unspecific products. Oligonucleotide synthesis companies recognize an asterisk between the selected bases as the symbol for ordering a phosphorothioate bond. 

Example P5 primer with 2 phosphorothioate bonds at the 3' end: 

5'- AATGATACGGCGACCACC*G*A -3'