Aperçu du produit
These kits are designed for the rapid preparation of plasmid DNA from Escherichia coli.
Plasmid MiniPrep Kit
This kit is designed for the rapid preparation of plasmid DNA from small cultures of Escherichia coli using convenient spin columns. The plasmid DNA is preferentially purified from other cellular components such as genomic DNA and RNA. This kit is able to purify plasmids up to 13,000 bp in size, and the typical purification yield is up to 20 μg from 1.5 mL of bacterial culture. Purified DNA is of excellent quality for transformation, restriction enzyme digestion, sequencing and more. Also available in a 96-well format.
Plasmid MiniPrep Kit (Magnetic Bead System and High Throughput Magnetic Bead System)
Norgen's Plasmid MiniPrep Kit (Magnetic Bead System) is designed for the rapid preparation of plasmid DNA from small batch cultures of Escherichia coli. Norgen’s magnetic beads bind DNA under optimized salt concentrations and release the bound DNA under low salt and slightly alkali conditions. The plasmid DNA is preferentially purified from other cellular components such as genomic DNA and RNA. The purified plasmids are fully digestible with all restriction enzymes tested, and are completely compatible with real-time PCR and NGS.
Norgen's Plasmid MiniPrep Kit (Magnetic Bead System) is also available in a 96-well (HT) format for high throughput applications. Purification with the 96-well plates can be integrated with a robotic automation system.
Plasmid MaxiPrep Kit
This kit is designed for the rapid spin column preparation of plasmid DNA from up to 100 mL of Escherichia coli cultures. The kit allows for the isolation of plasmid DNA with final endotoxin levels of 0.1 EU/µg of DNA or less. The kit is able to purify plasmids up to 13,000 bp in size, and typical yields from a 100 mL culture for a high copy number plasmid are between 0.4 and 1.0 mg. The purified DNA is fully digestible with all restriction enzymes tested, and is completely compatible with manual or automated sequencing to achieve 95-100% accuracy.
Détails
Supporting Data
Figure 1. High DNA Yield from Small Cultures. Plasmid DNA from 1 mL overnight bacterial cultures was prepared with Norgen’s Plasmid MiniPrep Kit and compared with two other commercial kits. The purifications were performed in triplicate, and the average yield for each kit is shown in the graph. The quantification of the DNA yield was performed by resolving 5 µL of the 50 µL of eluted DNA on a 1X TAE, 0.9% agarose gel followed by densitometry. Norgen shows a significantly higher average yield than the other 2 competitor’s kits.
Figure 2. High-Accuracy Sequencing. One microgram of plasmid DNA purified by Norgen’s Plasmid Miniprep DNA Kit was used as a template in an Applied Biosystem DNA Sequencer. The result showed an accuracy of >99% over a 950 bp contiguous sequence.
Figure 3. High Quality of Plasmid DNA. Resolution of plasmid DNA isolated from E. coli (1 mL) using Norgen’s Plasmid MiniPrep Kit (Magnetic Bead System). For evaluation, 10 µL from 50 µL of elution were digested with the restriction enzyme, EcoR I and run on 1X TAE 1.3% agarose gel. All plasmid DNA isolated were digestible, indicating the high quality of plasmid DNA. Marker = Norgen’s HighRanger DNA Ladder.
Figure 4. High DNA Yield from 150 mL Cultures. Plasmid DNA from 150 mL bacterial cultures grown overnight was prepared with Norgen’s Plasmid DNA MaxiPrep Kit and compared with two other commercial kits. The purifications were performed in triplicate, and the average yield for each kit is shown in the graph. The quantification of the DNA yield was performed by resolving 5 µL of the 2 mL of eluted DNA on a 1X TAE, 0.9% agarose gel followed by densitometry. Norgen’s kit shows a significantly higher average yield than the other 2 competitor’s kits.
Figure 5. Full Compatibility with Digests. DNA isolated with Norgen’s Plasmid DNA MaxiPrep Kit is easily digestible, often requiring less than 1 hour for full digestion. One microgram of a 9,309 bp plasmid purified by Norgen’s DNA MaxiPrep Kit was digested for one hour at 37oC in a 20 µL reaction with 2 units of BamHI (Lane 2), HindIII (Lane 3), and SmaI (Lane 4). The entire reaction was loaded on a 1X TAE, 0.9% agarose gel. Lane 1 is uncut plasmid, and Lane M is the Norgen UltraRanger 1kb DNA Ladder
Kit Specifications
|
|
Column Binding Capacity |
25 µg
|
Size of Plasmids Purified |
Up to 13,000 bp
|
Average Yield from 1.5 mL of Culture |
Up to 20 µg
|
Time to Complete 10 Purifications |
30 minutes
|
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the of shipment. The RNase should be stored at -20°C upon arrival. The Resuspension Solution AZ should be stored at 4oC upon addition of RNase enzyme.
Component | Cat. 13300 (50 preps) | Cat. 46400 (250 preps) | Cat. 46500 (4 preps) | Cat. 46600 (20 preps) | Cat. 60300 (50 preps) | Cat. 63000 (192 preps) |
---|---|---|---|---|---|---|
Resuspension Solution AZ | 12 mL | 60 mL | 20 mL | 100 mL | 12 mL | 2 x 20 mL |
Lysis Buffer N | 40 mL | 80 mL | 40 mL | 2 x 80 mL | 40 mL | 2 x 40 mL |
Buffer TN | 20 mL | 130 mL | 55 mL | 2 x 130 mL | 20 mL | 1 x 55 mL 1 x 20 mL |
Wash Solution E | 12 mL | 2 x 18 mL | - | - | - | - |
Elution Buffer K | 8 mL | 30 mL | - | - | 8 mL | 2 x 8 mL |
Wash Solution J | - | - | 25 mL | 3 x 25 mL | - | - |
Elution Buffer J | - | - | 24 mL | 120 mL | - | - |
RNase A | 1 vial | 1 vial | 1 vial | 1 vial | 1 vial | 1 vial |
Magnetic Bead Suspension | - | - | - | - | 1 x 1.1 mL | 4 x 1.1 mL |
Spin Columns | 50 | 250 | - | - | - | - |
Collection Tubes | 50 | 250 | - | - | - | - |
DNA Maxi Spin Columns with Collection Tubes (Clear ring in column) | - | - | 4 | 20 | - | - |
Maxi Spin Filter Columns with Collection Tubes (Grey ring in column) | - | - | 4 | 20 | - | - |
96-Well Plate | - | - | - | - | - | 2 |
Elution Tubes (1.7 mL) | 50 | 250 | - | - | 50 | - |
Elution Tubes (50 mL) | - | - | 4 | 20 | ||
96-Well Elution Plate | - | - | - | - | - | 2 |
Adhesive Tape | - | - | - | - | - | 2 |
Product Insert | 1 | 1 | 1 | 1 | 1 | 1 |
Documentation
(46400) Plasmid MiniPrep Kit - (250 prep)
(46500) Plasmid DNA MaxiPrep Kit - Protocol (4 prep)
(46600) Plasmid DNA MaxiPrep Kit - Protocol (20 prep)
(60300) Plasmid MiniPrep Kit (Magnetic Bead System) - Protocol (50 prep)
(63000) Plasmid MiniPrep 96-Well Kit (High Throughput Magnetic Bead System) - Protocol (2 x 96-well)