YALEPIC® Endotoxin Free Plasmid DNA Midi Isolation Kit (5-15ml)

This kit is standardly suitable for 5-15 mL of overnight E. coli culture. For high-copy plasmids, 5 mL of culture can meet the 100 microgram yield requirement. For low-copy plasmids or plasmids with large backbone (>10 kb), it is recommended to use 10-15 mL of culture to achieve more ideal yield.

The endotoxin carryover of plasmid DNA extracted with this kit is <=1 EU/microgram, meeting transfection-grade plasmid standards. This low endotoxin level fully meets the transfection requirements of various common cell lines such as HEK293, CHO, and HeLa, and is compatible with various transfection reagents such as Lipofectamine 3000, PEI, and the YALEPIC LFDR series, significantly reducing the negative impact of endotoxin on transfection efficiency and cell viability.

From culture collection to obtaining purified plasmid DNA, the entire operation workflow takes approximately 25 minutes. Optimized procedural steps and an efficient binding membrane system greatly shorten the time cost of traditional plasmid extraction. The visual blue indicator also helps users quickly determine whether lysis and neutralization are complete, further reducing operation waiting and re-verification time.

The blue indicator is a pre-mixed component of this kit. Its main function is to visually indicate whether lysis and neutralization steps are complete through color change – the solution turns blue when lysis is sufficient, and the blue color fades or becomes colorless/light when neutralization is complete. This design effectively avoids reduced plasmid yield and purity issues caused by insufficient lysis or over-neutralization, making it especially suitable for novice users or high-throughput operation scenarios.

Plasmid DNA extracted with this kit has extremely low endotoxin carryover and minimal salt carryover, and is perfectly compatible with the following downstream experiments: Cell transfection (recommended, suitable for various endotoxin-sensitive cell lines); Restriction enzyme digestion; PCR amplification and qPCR quantification; Sanger sequencing and NGS library construction; In vitro transcription (e.g., mRNA synthesis template); Competent cell transformation; Ligation and cloning.

This kit employs a dual-effect endotoxin removal mechanism: First, large-molecule endotoxin complexes are physically removed through a filtration column after cell lysis. Then, a specialized endotoxin removal solution system performs chemical clearance. The silica membrane specifically binds plasmid DNA while effectively excluding impurities such as endotoxin, protein, and genomic DNA, ultimately yielding high-purity endotoxin-free plasmid DNA.

Yes, but it is recommended to appropriately increase the culture volume. For low-copy plasmids (e.g., pACYC and pSC101 backbones) or large plasmids >10 kb, 10-15 mL of overnight culture is recommended. The lysis time can also be appropriately extended (approximately 1-2 minutes) to ensure complete cell lysis. During elution, using Elution Buffer preheated to 65 degrees C helps improve the elution efficiency of large plasmids.

Compared with imported brands, the YALEPIC kit offers equivalent performance quality at a more cost-effective price. Compared with domestic brands, this kit delivers equivalent yield (100 microgram) and endotoxin standard (<=1 EU/microgram) for the same 5-15 mL culture processing volume, while the visual blue indicator design is a core differentiating highlight. Localized production and fast logistics response also provide domestic users with higher service efficiency.

This kit supports ambient-temperature shipping (10-30 degrees C). Upon arrival, it can be stored directly at room temperature without refrigeration. Do not freeze any components, as freezing may cause reagent performance degradation. The product shelf life is 12 months (subject to the product label). Use within the shelf life and store properly after opening according to the instructions.

Common causes and solutions are as follows: High endotoxin – incomplete neutralization after lysis, buffer not mixed until no localized shadow remains. Thoroughly mix by inverting to ensure complete neutralization before filtration. Insufficient yield – insufficient culture volume or poor bacterial growth state. Use freshly overnight-cultured bacteria with OD600 recommended at 2.0-4.0. Poor purity – insufficient wash steps. Wash twice as described in the manual and ensure wash buffer has completely drained before elution. Difficult elution of large plasmids – preheat elution buffer to 65 degrees C before use, which significantly improves elution efficiency.