Endotoxin Free Midi Kit (PIFE25)

The Presto™ Midi Plasmid Kit Endotoxin Free was designed for Plasmid DNA Purification and endotoxin removal from 50-150 ml of cultured bacterial cells using an efficient plasmid midiprep system. For processing smaller volumes, the Presto™ Mini Plasmid Kit is also available. Both plasmid kits include TrueBlue Lysis Buffer (an optional color indicator) to prevent common handling errors, while ensuring efficient cell lysis and neutralization. The Presto™ Midi Plasmid Kit Endotoxin Free uses a modified alkaline lysis method and RNase treatment  to obtain clear cell lysate with minimal genomic DNA/RNA contaminants. Using an efficient gravity-flow procedure, plasmid DNA in the crude lysate is bound by the plasmid midi column and the contaminants are removed with PMC Buffer. The purified plasmid DNA is eluted by a high salt buffer and then precipitated with Isopropanol for desalting. The entire procedure can be completed in 80 minutes without ultracentrifuges, HPLC or other toxic reagents.   

Advantages (Cat. # PIFE25)

Presto™ Midi Plasmid Kit Endotoxin Free Functional Test Data

Brand Test 260/280 260/230 Yield Average Geneaid (Lane 1, 2) 50 ml 1.85 2.25 273.00 µg 265.90 µg  50 ml 1.87 2.23 258.80 µg 100 ml 1.87 2.30 373.20 µg 391.20 µg 100 ml 1.87 2.14 409.20 µg Competitor (Lane 3, 4) 50 ml 1.85 2.26 121.10 µg 131.30 µg 50 ml 1.85 2.29 141.50 µg 100 ml 1.86 2.32 285.80 µg 287.85 µg 100 ml 1.87 2.33 289.90 µg

Figure 1. Plasmid DNA was extracted using both the Presto™ Midi Plasmid Kit Endotoxin Free (lane 1,2) and the equivalent competitor's plasmid midi kit (lane 3,4). The purified supercoiled plasmid DNA [50 ml and 100 ml overnight E. coli (DH5α) culture, containing a 3 kb plasmid pBluescript (A600 > 2 U/ml, OD600 = 3.8)], was used in EcoRI digestion and analyzed by electrophoresis on a 1% agarose gel. M = Geneaid 1 Kb DNA Ladder, Lane 1: Presto™ Midi Plasmid Kit Endotoxin Free (50 ml), Lane 2: Presto™ Midi Plasmid Kit Endotoxin Free (100 ml), Lane 3: Equivalent Competitor Kit (50 ml), Lane 4: Equivalent Competitor Kit (100 ml)

Column Types Included With the Presto™ Midi Plasmid Kit Endotoxin Free

Filter Column
	Figure 2. This centrifuge midi plasmid spin column is used to filter the neutralized colorless mixture and remove contaminants to facilitate DNA binding to the white membrane of the Plasmid Midi Column.
Plasmid Midi Column
	Figure 3. This gravity flow plasmid midi anion exchange resin column is used for binding, washing and eluting plasmid DNA.

TrueBlue Lysis Buffer Optional Color Indicator

Cell Lysis		Neutralization		Figure 4. TrueBlue Lysis Buffer is an optional color indicator included with this Plasmid Kit to eliminate common handling errors and ensure efficient cell lysis and neutralization each and every time.
Insufficient Mixing	Correct Mixing	Insufficient Mixing	Correct Mixing

Plasmid DNA Midi and Maxi Kits Specifications

Specifications	Presto™ Midi Plasmid Kit	Presto™ Midi Plasmid Kit Endotoxin Free	Geneaid™ Midi Plasmid Kit	Geneaid™ Midi Plasmid Kit Endotoxin Free	Geneaid™ Maxi Plasmid Kit	Geneaid™ Maxi Plasmid Kit Endotoxin Free
Cat. #	PIF025	PIFE25	PI025	PIE25	PM010/25	PME10/25
Format	Anion-Exchange Gravity Flow	Anion- Exchange Gravity Flow	Anion-Exchange Gravity Flow	Anion- Exchange Gravity Flow	Anion-Exchange Gravity Flow	Anion- Exchange Gravity Flow
Binding Capacity	350 µg	350 µg	350 µg	350 µg	1.0 mg	1.0 mg
Filter Column	Yes	Yes	N/A	N/A	N/A	N/A
Endotoxin Level	<0.15 EU/µg	<0.10 EU/µg	<0.15 EU/µg	<0.10 EU/µg	<0.15 EU/µg	<0.10 EU/µg
Culture Input	50-150 ml	50-150 ml	50-300 ml	50-300 ml	200-1600 ml	200-1600 ml
Plasmid Size	1-20 kb	1-20 kb	1-20 kb	1-20 kb	1-20 kb	1-20 kb
Typical Yield	350 µg	350 µg	350 µg	350 µg	1.0 mg	1.0 mg
Elution Volume	500 µl-2 ml	500 µl-2 ml	500 µl-2 ml	500 µl-2 ml	500 µl-2 ml	500 µl-2 ml
Operation Time	<80 minutes	<80 minutes	<80 minutes	<80 minutes	<100 minutes	<100 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