最新产品
Direct Lysis Plasmid96 DNA Purification Kit
Direct Lysis Plasmid96 DNA Purification Kit
货号
PLS84596
规格
5 x 96 well preps
存储温度
Room Temperature
产品详情
- 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.
Benefits
- 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).
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Overnight culture: Add lysis buffer; mix. Add neutralization buffer; mix. |
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No cell harvesting. No use of deep-well culture plate. |
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Clear the lysate: |
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Precipitate the DNA: Centrifuge, 10 min. Wash with 70% EtOH, 5 min. Resuspend the DNA. |
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Figure 1. Overview of the Direct Lysis Plasmid96 purification process. |
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| 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 |
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| Direct Lysis Plasmid96 DNA Purification Kit | |
| PLS84596 | 5 x 96-well preps |
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Contents: Five culture plates, five 96-well fritted filter plates, five collection plates, plate sealers, gas-permeable sealers, and reagents for five 96-well preps. |
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