DNA & RNA PLASMID PURIFICATION

Plasmid DNA plays a pivotal role in molecular biology research, but obtaining it from prokaryotic cultures can be tricky. It needs to be separated from genomic DNA in a complex lysis and purification procedure. We supply products that allow plasmid DNA to be obtained from prokaryotic cultures in a way that preserves higher amounts of plasmid DNA than can be found in pre-packaged samples and at a lower cost.    

Plasmid DNA is used to express recombinant proteins in various host organisms during molecular biology and protein research. Progress in cloning technology has led to the need for numerous samples of small cultures of plasmid DNA. Small-scale preparation of plasmid DNA requires parallel purifications using semi-automated or fully-automated handling.

Lysing is the first step in purifying plasmids from within the cultured cells. Once the DNA has successfully been extracted, it must be clarified in order to remove cellular debris. Many manufacturers have created kits for preparing large and small samples of plasmid DNA for research use. These kits are expensive and inefficient. They include single spin tubes in addition to 96-well plate formats.

When using a single spin tube for plasmid purification, centrifugation is usually used to clarify the DNA, giving rise to several complications. Spin times are long, the rotor capacity is insufficient for single tube processing, and the fluffy pellets used during the process are apt to trap a significant amount of the plasmid DNA due to sedimentation.

Lysate Clearance Quick Tech Note

We present a lysate sample preparation procedure using the AcroPrep™ Advance filter plate for lysate clearance that effectively removes unwanted cellular debris from samples.

The AcroPrep Advance 96-well filter plates provide the ability to use equipment for complete automation of the DNA purification process. The filter plates are manufactured out of material that allows for optimal DNA binding through the use of chaotropic salts, and the new well geometry ensures higher recovery of DNA. Contaminants are then conveniently washed away. The filter plates allow for centrifugation or vacuum filtration, which can be automated and increases DNA recovery through an additional wash step. The filter plates also contain a prefilter for lysate clarification, which efficiently collects the clarified lysate into a DNA binding plate.

Additional methods of purification can include a chromatography step. We offer an innovative chromatography membrane which can provide higher dynamic binding capacity compared with traditional resin based solutions.

Our Mustang® chromatography units facilitate the removal of host cell proteins and work to purify plasmid DNA. They are easy to use and disposable, creating an optimally sterile environment for purification using ion exchange chromatography.

AcroPrep Advance MWCO 96-well filter plates can be used to concentrate DNA if pooling multiple fractions before running downstream analytics is required. 

Typically, further downstream applications can involve PCR and the requirement to purify unincorporated primers and dNTPS away from the PCR product. We supply a range of AcroPrep Advance MWCO 96-well filter plates and centrifugation products that can assist with such applications.