Plasmid ezFilter Maxi Kit (10 Preps)

Plasmid ezFilter Maxi Kit (10 Preps)

Cat. No.: BMG-PD1512-01
Size: 10 Preps


Description

Key to the kit is our proprietary DNA binding systems that allow 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-PD1512-01	BMG-PD1512-2
Maxi Column	10	25
Syringe Filters	10	25
Buffer A1	120 mL	260 mL
Buffer B1	120 mL	260 mL
Buffer C1	140 mL	320 mL
RNase A	12 mg	25 mg
Elution Buffer	15 mL	50 mL
EndoClean Buffer	N/A	N/A

Caution: Buffer C1 contains acetic acid, wear gloves and protective eyewear when handling.


Important:

• RNase A: Spin down RNase A vial briefly. Add the RNase A solution to buffer A1 and mix well before use.
• Buffer B1 precipitates below room temperature, it is critical to warm up the buffer at 50°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

• 70% ethanol and 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 capable of generating 200-300 mbar.
• 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	Copy Numbers	Expected Yield (µg /200 mL)
pSC101	pSC101	5	10-15
pACYC	P15A	10-12	20-25
pSuperCos	pMB1	10-20	20-40
pBR322	pMB1	15-20	30-40
pGEMR	Muted pMB1	300-400	400-500
pBluescriptR	ColE1	300-500	400-600
pUC	Muted pMB1	500-700	600-1200

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, 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 OD₆₀₀ 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 (OD₆₀₀). 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.