NUCLEIC ACIDS EXTRACTION
FavorPrep™ Plasmid DNA Extraction Mini Kit
|
Description
The FavorPrep™ Plasmid Extraction Kit provides a rapid, phenol-free method for the extraction of high-purity plasmid DNA from bacterial cultures such as E. coli , which bacteria is pellet, lysed, and then neutralized. The extracted DNA can be used in a variety of applications such as PCR, cloning, sequencing, in vitro transcription, and labeling. Also, as a column-type tube is utilized in the purification process, extraction is carried out in three simple steps of binding/washing/elution. Once bound, the DNA is washed and then eluted from the column, ready for use.
Features
• For high yields of plasmid DNA-up to 30£gg from 1~5ml overnight cultures.
• Effective purification of DNA fragments ranging from 100bp to 12+kb.
• No need for messy resin slurries, extracting with phenol, or concentrating via alcohol precipitation.
• Superior purity-DNA yields quality sequence data using automated or manual methods.
• Optimized buffers are included for maximum DNA purity and yield.
• Versatile protocol-works with all neutral gel buffers and both conventional and low-melt agarose gel.
Format
Spin Columns
Operation
Centrifuge/vacuum
Binding Capacity
Up to 30 £gg
Expectant Yield
10-20 £gg for high-copy plasmid
1-5£gg for low-copy plasmid
Time Required
Under 30 minutes
DNA Sequencing Analysis
Sequencing of plasmid purified with FavorPrep Plasmid Extraction Mini Kit. The sequence was analyzed with BigDye Terminator chemistry on ABI 3700
Applications
• Fluorescent or radioactive Sequencing
• Ligation
• Restriction enzyme digestion
• Ligation and Transformation
• Library screening
Agarose Gel Analysis Of Plasmids Purified
| M1 |
1 |
2 |
3 |
4 |
5 |
6 |
M2 |
 |
Restriction analysis of 4 plasmids extracted by the FavorPrep? Plasmid DNA Extration Kit. Digestion with EcoRI.
M: 1kb DNA ladder (100bp-12,000bp)
1: pBluescript II
2. pUC 18
3: pBR 322
4: pGem with 1Kb insert
5: pET 43.1 with 2.7Kb insert
6: cosmid (50Kb)
M1: 1Kb DNA Ladder
M2: Lambda-Hind III
Procedure
The modified alkaline lysis method and RNase treatment are used for cteating cleared cell lysate with minimal genomic DNA and RNA contaminants. In the presence of a chaotropic salt, the plasmid DNA in the lysate binds to the glass fiber matrix in the spin column. The purified plasmid DNA is eluted by a low salt elution buffer or water. The procedure doesn't require DNA phenol extraction or alcohol precipitation.
Bacterial Cells
 |
Cell Harvesting
|
Resuspension
Lysis
Neutralization |
DNA Binding |
Wash |
Elution |
Storage Conditions
The Plasmid DNA Extraction Kit can be stored at room temperature (15 -25 ¢J ). After adding RNase A, PDE Buffer1, should be stored at 4 ¢J and is stable for six months. Other buffers and columns can be stored dry for up to 1 year at room temperature (15 -25 ¢J ).
| Ordering Information |
| Cat. No. |
Product Name |
Size |
Kit Contents |
Store at |
FAPDE 001 |
FavorPrep™ Plasmid DNA Extraction Kit |
100 preps. |
1. FAPD 1 Buffer
2. FAPD 2 Buffer
3. FAPD3 Buffer
4. Wash Buffer1
5. Wash Buffer (conc.)
6. Elution Buffer
7. RNase (50 mg/ml)
8. PDE Column
9. Collec tion Tube
|
25ml
25ml
45ml
45ml
20ml+80ml
6ml
50 £gl
100 preps
100 tubes
|
PDE Buffer with RNaseA at 4 ¢J , others at room temperature.
(15 -25 ¢J ) .
|
FAPDE 001-1 |
FavorPrep™ Plasmid DNA Extraction Kit |
300 preps. |
1. PDE Buffer 1
2. PDE Buffer 2
3. PED Buffer 3
4. Wash Buffer 1
5. Wash Buffer (conc.)
6. Elution Buffer
7. RNase (100 mg/ml)
8. PDE Column
9. Collection Tube |
65ml
75ml
100ml
130ml
50ml
30ml
130£gl
300 preps
300 tubes
|
PDE Buffer with RNaseA at 4 ¢J , others at room temperature.
(15 -25 ¢J ) .
|
|
