Home FAQs Does sequence length determine dbDNA™ (doggybone™ DNA) yield?

FAQ: Does sequence length determine dbDNA™ (doggybone™ DNA) yield?

No. The EnClose™ Cell-free dbDNA Synthesis Kit (NEB #E9301) is specified to produce ≥10 µg dbDNA per 100 µl reaction using the dbDNA Vector as the input, but the yield may vary depending on the sequence of interest (SOI). For most constructs (~1.5–12 kb), sequence length does not directly correlate with yield. Yields are broadly distributed across this range. However, very short (~500 bp) or very long (~20 kb) constructs are more likely to show reduced yields. 

Final dbDNA yields are more likely to be affected by sequence content, not length. Rolling circle amplification (RCA) can be inherently variable. While design choices can help, yields may be higher or lower than expected for a given sequence. Features such as long homopolymers, strong terminators, extensive secondary structure, repetitive elements, or ITRs can reduce yield.

However, once a construct’s yield is known, reaction-to-reaction (or batch‑to‑batch) variability is typically low.