Introduction
The yield of plasmid DNA depends on the origin of the replication and the size of the plasmid. The protocols are optimized for high copy number plasmid purification. For low copy number plasmids, both the culture volume and the buffer volume need to be scaled up 3 to 5 times. Please contact our customer service for further information and reference the table below for the commonly used plasmids.
Specifications
|
Plasmid |
Origin |
High copy |
Low copy |
|
pACYC |
P15A |
|
10-12 |
|
pSC101 |
pSC101 |
|
5 |
|
pSuperCos |
pMB1 |
|
10-20 |
|
pBR322 |
pMB1 |
|
15-20 |
|
pUC |
Muted pMB1 |
500-700 |
|
|
pGEMR |
Muted pMB1 |
300-400 |
|
|
pBluescriptR |
ColE1 |
300-500 |
|
Materials supplied by user
- Centrifuge with swing-bucket rotor (4,000 x g).
- Please disinfect 96-well 2.2 mL plates before use.
- Vacuum pump capable of achieving 300-400 mbar.
- Standard vacuum manifold.
- Oven or incubator preset to 70 oC.
Storage and Stability
All components are guaranteed for 24 months from the date of purchase. The Buffer A1/RNase A should be stored at 4oC.
Kit Contents
| Catalog# |
PD1812-S |
PD1812-01 |
PD1812-02 |
| Preps |
1 |
4 |
20 |
| 96- Well 2.2 mLPlate |
1 |
4 |
20 |
| 96- Well 1.6 mLPlate（Can be reused） |
1 |
4 |
20 |
| 96-Well DNA Plate |
1 |
4 |
20 |
| 96-Well Lysate Clearance Plate |
1 |
4 |
20 |
| 96-Well Collection Plate |
2 |
5 |
24 |
| Ventilating Film |
1 |
4 |
20 |
| Sealing Film |
4 |
16 |
80 |
| Buffer A1 |
30 mL(Add RNase A before use) |
110 mL(Add RNase A before use) |
2 x 300 mL |
| Buffer B1 |
30 mL(Keep tightly capped after use) |
110 mL(Keep tightly capped after use) |
2 x 300 mL |
| Buffer N1 |
40 mL(Contain chaotropic salts) |
160 mL(Contain chaotropic salts) |
2 x 400 mL |
| DNA Wash Buffer |
50 mL(Add 200 mL ethanol before use) |
2×100 mL(Add 400 mL ethanol before use) |
7 x125 mL(Add 500 mL ethanol before use) |
| Buffer KB |
55 mL |
240 mL |
3 x 400 mL |
| Elution Buffer |
25 mL |
100 mL |
450 mL |
| RNase A(20mg/mL) |
160 μL |
600 μL |
2 x 1.5 mL |
| Use Manual |
1 |
1 |
1 |
Before Starting
Exam this handbook and get familiar with each step. Prepare all components and have the necessary materials ready.
- Briefly spin down the RNase A vial and add the RNase A to Buffer A1.
- Dilute DNA Wash Buffer as follows:
PD1812-00: Add 200 mL 96-100% ethanol to each bottle before use.
PD1812-01: Add 400mL 96-100% ethanol to each bottle before use.
PD1812-02: Add 500mL 96-100% ethanol to each bottle before use.
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