Introduction
EZgeneTM yeast plasmid kit is designed for rapid and reliable isolation of high-quality plasmid DNA from yeast cultures. Utilizing the reversible nucleic acid-binding properties of our matrix, the plasmid DNA is bound to the matrix while proteins and other unwanted impurities are eliminated by wash buffer. Pure DNA is then eluted. Purified DNA can be directly used in downstream applications such as PCR, restriction digestion, and Southern Blot.
The Yeast Plasmid Mini Kit combines the power of spin column technology with the lyticase, glass beads and alkaline-SDS lysis of yeast cells to yield high quality plasmid DNA in less than 1 hour. The mini spin columns facilitate the binding, washing, and elution steps, thus enabling multiple samples to be processed simultaneously. The actual plasmid yields depend on copy numbers, yeast strain, and conditions of growth. Because of low copy numbers, the maximum yield from 5 mL yeast culture is around 1 µg.
This protocol has been successfully used to isolate autonomous plasmids from S. cerevisiae. As a modified alkaline lysis procedure, genomic DNA is virtually eliminated from the preparation. Note that all centrifugation steps should be carried out at room temperature.
Storage and Stability
All EZgeneTM Yeast Plasmid Mini Kit components are guaranteed for at least 12 months from the date of purchase when stored as follows: Buffer YP I/RNase A mix at 4oC, Lyticase solution at -20oC, all other components at room temperature.
Capacity
The binding capacity for the mini column is 40 µg of plasmid DNA.
Kit Components
|
Catalog# |
YD1291-S |
YD1291-01 |
YD1291-02 |
|
Preps |
1 |
4 |
20 |
|
96-Well DNA Plate |
1 |
4 |
20 |
|
Deep Well Plate |
1 |
4 |
20 |
|
Collection Plate (300μL) |
1 |
4 |
20 |
|
96-Well Lysate Clearance Plate |
1 |
4 |
20 |
|
Buffer YP I |
30 mL |
110 mL |
550 mL |
|
Buffer YP II |
30 mL |
110 mL |
550 mL |
|
Buffer YPIII |
40 mL |
160 mL |
800 mL |
|
Lyticase |
300 U |
12,000 U |
60,000 U |
|
Buffer SE |
50 mL |
200 mL |
1000 mL |
|
Glass Beads |
5.4 g |
22 g |
110 g |
|
DNA Wash Buffer |
50 mL (Add 150 mL ethanol before use) |
2×100 mL (Add 300 mL ethanol before use) |
7×125 mL (Add 325 mL ethanol before use) |
|
Elution Buffer |
15 mL |
60 mL |
300 mL |
|
RNase A Concentrate |
140 μL |
560 μL |
2,800 μL |
|
Instruction Booklet |
1 |
1 |
1 |
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