Home FAQs What yields can I expect from the EnClose™ Cell-free dbDNA™ Synthesis Kit (NEB #E9301)

FAQ: What yields can I expect from the EnClose™ Cell-free dbDNA™ Synthesis Kit (NEB #E9301)

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.


EnClose Cell-free dbDNA Synthesis Kit provides high dbDNA yield across constructs

Gel depicting high dbDNA yield for various input plasmid sizes 
The dbDNA workflow supports a range of input plasmid and product sizes, delivering > 90% yield of the desired dbDNA; yield variation depends on sequence of interest (SOI) size and features. Product yields were quantified using an Unchained Labs® Lunatic® spectrophotometer. 50 ng of DNA sample was loaded into each lane of a BioAnalyzer® 12000 to determine purity of correct dbDNA product.