Plasmid Maxi Kit (2 Preps)
Cat. No.: BMG-PD1511-00
Size: 2 Preps
Description
Key to the kit is our proprietary DNA binding system that allows the highly efficient binding of DNA to our ezBindTM matrix while proteins and other contaminates are removed under certain optimal conditions. Nucleic acids are easily eluted with sterile water or TE buffer. Unlike all other rivals, Biomiga’s patented plasmid purification kit has no guanidine salt in the buffer, the purified DNA is guanidine/ion exchange resin residues free which enable the high performance of downstream applications such as transfection, restriction mapping, library screening, sequencing, as well as gene therapy and genetic vaccinations.
Storage and Stability
Buffer A1 should be stored at 4°C once RNase A is added. All other materials can be stored at room temperature. The Guaranteed shelf life is 12 months from the date of purchase.
Kit Contents
| Catalog# |
BMG-PD1511-01 |
BMG-PD1511-2 |
| Preps |
10 |
25 |
| Maxi Column |
10 |
25 |
| Buffer A1 |
120 mL |
260 mL |
| Buffer B1 |
120 mL |
260 mL |
| Buffer C1 |
140 mL |
320 mL |
| DNA Wash Buffer |
50 mL |
2x50 mL |
| EndoClean Buffer |
10 mL |
25 mL |
| Elution Buffer |
25 mL |
60 mg |
| RNase A |
420 µL |
900 µL |
Important:
- RNase A: Spin down RNase A vial briefly. Add the RNase A solution to buffer A1 and mix well before use.
- Add 200 mL 100% ethanol to each DNA Wash Buffer.
- Buffer B1 precipitates below room temperature, it is critical to warm up the buffer at 37°C to dissolve the precipitates before use.
- Keep the cap tightly closed for buffer B1 after use.
- Ensure the availability of centrifuge capable of 13,000 rpm.
- Carry out all centrifugations at room temperature.
Materials required but not supplied
- 100% ethanol.
- Miniprep: 1.5 mL eppendorf tubes and centrifuge.
- Midi/maxiprep: high speed centrifuge, 30 mL high speed centrifuge tubes, 15 /50 ml tubes, and or vacuum manifold.
- Megaprep: Vaccum system, 500 mL bottle (Corning# 430282) or 1,000 mL bottle (#430518) or equivalent.
Important Notes
Copy numbers: 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. reference the table below for the commonly used plasmids
| 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 |
|
Host Strains: The strains used for propagating plasmid have significant influence on yield. Host strains such as Top 10, DH5a, and C600 yield high-quality plasmid DNA.
endA+ strains such as JM101, JM109, JM110, HB101, TG1 and their derivatives, normally have low plasmid yield due to either endogenous endonucleases or high carbohydrates released during lysis. We recommend transform plasmid to an
endA- strain if the yield is not satisfactory. For purifying plasmid DNA from
endA+ strains, we recommend use product number PD1713.
Optimal cell mass (OD600 x mL of Culture): This procedure is designed for isolating plasmid grown in standard LB medium (Luria Bertani) for 12 -16 hours to a density of OD
600 2.0 to 3.0. If rich medium such as TB or 2xYT are used, make sure the cell density doesn’t exceed 3.0 (OD
600). A high ratio of biomass over lysis buffer results in low DNA yield and purity. The maxi column has an optimal biomass of 400-500. For example, if the OD600 is 2.5, the optimal culture volume should be 150 to 200 mL.
