Direct Lysis Plasmid96 DNA Purification Kit

Direct Lysis Plasmid96 DNA Purification Kit




5 x 96 well preps


Room Temperature


  • Rapid, direct-lysis preparation of plasmid DNA for sequencing in a 96-well format.

The Direct Lysis Plasmid96 Kit isolates plasmid DNA by direct lysis of a 200-µl overnight culture, eliminating the need for centrifugation of the culture and resuspension of the cell pellet (Fig. 1). The kit is designed for rapid preparation of sequencing templates in a 96-well format from either high-copy or single-copy plasmids sized up to 40 kb (Figs. 2 and 3). The kit uses a modified alkaline lysis procedure that incorporates a proprietary RNase Blend to remove the RNA efficiently. The typical yield from a 200-µl overnight culture of a high-copy plasmid vector is sufficient for up to 50 end-sequencing reactions. A copy-number-induced 40-kb plasmid will yield DNA for six to eight sequencing reactions.

The kit includes reagents and plastics for high-throughput DNA extraction, and is suitable for robotic automation procedures.


  • No cell harvesting or resuspension of the cell pellet.
  • Inoculation of culture in a 200-µl volume eliminates the need for 2-ml, deep-well growth plates.
  • Reduces the amount of media used to under 20 ml per plate compared to 150 ml with a standard 1.2-ml prep.
  • Low final DNA suspension volume (50 µl).
  • Protocol complete in as little as 25 minutes.
  • Perform ~50 end-sequencing reactions from a high-copy plasmid, with average read length of 700-900 bases (Fig. 3).
  • Low-profile filter-receiver plate block (H57 x L127 x W85 mm).
Overnight culture:
Add lysis buffer; mix.
Add neutralization buffer; mix.
Figure 1
No cell harvesting.
No use of deep-well
culture plate.

Clear the lysate:
Vacuum filtration or
centrifugation, 1 min.

Precipitate the DNA:
Centrifuge, 10 min.
Wash with 70% EtOH, 5 min.
Resuspend the DNA.
Figure 1. Overview of the Direct Lysis Plasmid96 purification process.


Figure 2. Examination of the quality of plasmid DNA. DNA was isolated from 200-µl overnight cultures of randomly picked clones from a shotgun library (2-6 kb) in a 96-well plate using the Direct Lysis Plasmid96 Kit. A 5-µl aliquot of each 50-µl prep was analyzed on a 1% agarose gel in 1X TAE buffer for 2 hours. Lane M, Supercoiled DNA Ladder; lanes 1-18, supercoiled plasmid DNA. Figure 3. Sequence analysis confirms plasmid DNA quality. Sequencing was performed using 2 µl of DNA isolated from a plasmid and M13 Forward primer, and was analyzed on an ABI® 3730 Analyzer in the UWBC DNA Sequencing and Synthesis Facility (Madison, WI).


Catalog No. Size

Direct Lysis Plasmid96 DNA Purification Kit
PLS84596 5 x 96-well preps
Five culture plates, five 96-well fritted filter plates, five collection plates, plate sealers, gas-permeable sealers, and reagents for five 96-well preps.