Large Plasmid Mini Kit (PDL100, PDL300)

The Large Plasmid DNA Extraction Kit was designed for plasmid and cosmid DNA purification from 1-4 ml of cultured bacterial cells. This plasmid kit is optimized for extracting 10-50 kb plasmid DNA using a miniprep spin column system. The Large Plasmid DNA Extraction Kit uses a modified alkaline lysis method and RNase treatment to obtain clear cell lysate with minimal genomic DNA and RNA contaminants. In the presence of chaotropic salt, plasmid DNA in the lysate binds to the glass fiber matrix of the plasmid spin column. Contaminants are removed with a Wash Buffer (containing ethanol) and the purified plasmid DNA is eluted by a low salt Elution Buffer or TE. Typical yields are 20-30 μg for high-copy number plasmid or 3-10 μg for low-copy number plasmid from 4 ml of cultured bacterial cells. DNA phenol extraction or alcohol precipitation is not required and the entire procedure can be completed within 15 minutes. The purified plasmid DNA is ready for use in Restriction Enzyme Digestion, Ligation, PCR, and sequencing reactions. 

Advantages (Cat. # PDL100, PDL300)  

• Purify 10-50 kb plasmid DNA within 15 minutes!
• High Yield: up to 30 μg of plasmid DNA or cosmid DNA
• Sample: 1-4 ml of bacterial cells (10-50 kb plasmid)
• Spin Columns: glass fiber membrane optimized for large plasmid DNA purification
• Elution Volume: 50-100 µl
• Storage: dry at room temperature (15-25ºC)

Applications

 

Restriction Enzyme Digestion, Library Screening, Ligation, PCR, Transformation and Sequencing Reactions

 

Components

• PDL1, PDL2, PDL3 Buffers
• WL1 Buffer
• Wash Buffer
• Elution Buffer (10 mM Tris-HCl, pH8.5 at 25°C)
• RNase A
• LP Columns
• 2 ml Collection Tubes

Quality Control

 

The quality of the Large Plasmid DNA Extraction Kit is tested on a lot-to-lot basis according to Geneaid's ISO-certified quality management system. 5 µl aliquots of a 100 µl eluate of purified super coiled plasmid DNA from a 4 ml overnight E. coli (DH5α) culture, containing a 17.4 kb low copy plasmid (OD600 = 3.5 U/ml) are analyzed by electrophoresis on a 0.8% agarose gel

 

Large Plasmid DNA Extraction Kit Functional Test Data



Figure 1. Plasmid DNA was extracted using the Large Plasmid DNA Extraction Kit. 5 µl aliquots of a 100 µl eluate of purified super coiled plasmid DNA from a 4 ml overnight E. coli (DH5α) culture, containing a 17.4 kb low copy plasmid pBluescript (OD600 = 3.5 U/ml) were used in EcoRI digestion and analyzed by electrophoresis on a 0.8% agarose gel. M = Geneaid 1 Kb DNA Ladder Test Conc. 260/280 260/230 Yield 1 87 ng/µl 1.90 2.16 8.7 µg 2 79 ng/µl 1.90 2.14 7.9 µg 3 86.4 ng/µl 1.89 2.17 8.6 µg

Plasmid DNA Kit Specifications

Specifications	Presto™ Endotoxin Free Mini Plasmid Kit	Presto™ Mini Plasmid Kit	High-Speed Plasmid Mini Kit	Large Plasmid DNA Extraction KIt	Presto™ 96 Well Plasmid Kit
Cat. #	PEH100	PDH100 PDH300	PD100 PD300	PDL100 PDL300	96PDC04/10 96PDV04/10
Format	Spin Column	Spin Column	Spin Column	Spin Column	96 Well Plate
Binding Capacity	40 µg	50 µg	30 µg	30 µg	35 µg
Culture Input	1-5 ml	1-7 ml	1-5 ml	1-4 ml	1-5 ml
Plasmid Size	1-15 kb	1-15 kb	1-15 kb	10-50 kb	1-15 kb
Typical Yield	up to 40 µg	up to 50 µg	10-30 µg	10-25 µg	20-35 µg/well
Elution Volume	30-100 µl	30-100 µl	30-100 µl	50-100 µl	100 µl
Operation Time	<60 minutes	<15 minutes	<15 minutes	<15 minutes	25 minutes

Related Plasmid DNA Purification Products

Product Name	RXNS	Catalogue Number
Presto™ Endotoxin Free Mini Plasmid Kit	100	PEH100
Presto™ Mini Plasmid Kit	100, 300	PDH100/300
High-Speed Plasmid Mini Kit	100, 300	PD100/300
Large Plasmid DNA Extraction KIt	100, 300	PDL100/300
High-Speed Plasmid Advance Kit (50-100 ml)	25	PA025
Presto™ Midi Plasmid Kit	25	PIF025
Presto™ Midi Plasmid Kit (Endotoxin Free)	25	PIFE25
Geneaid™ Midi Plasmid Kit	25	PI025
Geneaid™ Midi Plasmid Kit (Endotoxin Free)	25	PIE25
Presto™ Plasmid DNA Concentration Kit	25, 50, 100	PC0250/500/1000
Geneaid™ Maxi Plasmid Kit	10, 25	PM010/25
Geneaid™ Maxi Plasmid Kit (Endotoxin Free)	10, 25	PME10/25
Presto™ 96 Well Plasmid Kit	4 x 96, 10 x 96	96PDC04/10 96PDV04/10