Purification Made Easy with Microcentrifuge Spin Column Kits

Recent advancements in proteomic and genomic studies have increased the need for safe, high-throughput, and cost-effective equipment and processes for the isolation of microscale nucleic acids and purification of proteins.

A range of microcentrifuge spin column kits containing specifically designed membranes or filters are now commercially available for such applications.

Microcentrifuge Spin Column Kits

Plasmid DNA, genomic DNA, polymerase chain reaction (PCR) amplified DNA, and different types of RNA (mRNA, miRNA, total RNA and so on) from prokaryotic, eukaryotic and viral sources can be acquired using microcentrifuge spin column kits together with centrifugation.

These kits purify nucleic acids efficiently so that subsequent steps such as sequencing, RNA transcription, enzymatic restriction, transfection, qPCR and microinjection can be performed without interference from contaminants such as polysaccharides, proteins, metabolites and unwanted residual PCR products.

Microcentrifuge spin columns and kits are also available for the preparation of assays and the purification and concentration of proteins.

These columns enable small quantities of recombinant proteins to be purified quickly and simply and are based on size exclusion, affinity and other similar chromatographic principles. The filter-based columns are also used for protein concentration and buffer exchange.

Beckman Coulter’s Microfuge family of microcentrifuges offers rotors (Figure 1) that accommodate microcentrifuge spin columns. The Avanti high-performance centrifuges and the Allegra benchtop centrifuges deliver improved performance and versatility with additional combinations of labware and rotor.

Rotor that accommodates microcentrifuge spin columns.

Figure 1. Rotor that accommodates microcentrifuge spin columns.

GE Healthcare Life Sciences, QIAGEN, Bio-Rad, Promega, EMD Millipore, and Stratagene are leading providers of microcentrifuge spin column kits. These kits are available in the following formats:

  • Standard 96-well microtiter plates for high throughput
  • Single column or tubes with membranes or filters for use with small sample numbers
  • Strips of 8 (or 12) tips or columns

Square-well or deep-well plates can be used to support the 12 or 8 strips. Nucleic acids are captured onto the filter assembly using the filter membrane containing a large-pore, silica-based matrix/gel coated with a high density of anion exchange groups.

These assemblies are disposable and are usually spun from 100 x g to 30,000 x g for 15s to 30min, depending on the particular protocol used for the purification and separation procedure.

Quick Tips

The kit manufacturers’ instructions should be followed for optimum results.

  • Whenever small (microliter) volumes are used, the tube (or whole plate) should be tapped on the side or spun briefly at low speed (≥ 100 x g for 2 min) to make sure that droplets adhered to the tube’s side wall are collected towards the bottom, for thorough mixing and improved recovery.
  • The critical speed range for each centrifuge and rotor should be avoided during low-speed spins.
  • The centrifuge’s chamber and rotor should be cleaned after use to prevent damage from unintentional leaks and spills of solutions from open columns.

Table 1 lists the centrifuge rotor specifications:

Table 1. Centrifuge rotor specifications

Rotor

Maximum Labware Capability

Max. Speed (RPM/RCF x g)

Microfuge 16

FX121.5P

12, 1.5/2.2mL microcentrifuge tubes

14,800 / 15,183

FX241.5P

24, 1.5/2.2mL microcentrifuge tubes

14,800 / 16,163

Microfuge 20 Series

FA241.5 FA241.5P

24, 1.5/2.2mL microcentrifuge tubes

15,000 / 20,627

FA361.5

6, 1.5/2.0mL microcentrifuge tubes

15,000 / 20,124

Allegra X-30 Series

FX301.5

30, 1.5/2.0mL microcentrifuge tubes

16,000 / 28,672

F2402H

24, 1.5/2.0mL microcentrifuge tubes

18,000 / 29,756

S2096

2, 96 deep-well plates

3,000 / 1,109

Allegra X-12 Series

SX4750/A

4, 96 deep-well plates

3,750 / 2,885

Allegra X-14 Series

SX4750/A

4, 96 deep-well plates

4,000 / 3,283

Allegra X-15R

SX4750/A

4, 96 deep-well plates

4,450 / 4,060

Allegra 25R

TA-15-1.5

30, 1.5/2.0mL microcentrifuge tubes

15,000 / 25,160

S5700

2, 96 deep-well plates

5,700 / 6,130

Allegra 64R

F2402H

24, 1.5/2.0mL microcentrifuge tubes

26,000 / 61,970

F1202

12, 1.5/2.0mL microcentrifuge tubes

30,000 / 64,400

H6002

60, 1.5/1.8mL microcentrifuge tubes

12,200 / 12,400

F3602

36, 1.5/2.0mL microcentrifuge tubes

22,000 / 47,620

Avanti J Series

JA-18

30, 1.5mL microcentrifuge tubes

18,000 / 35,000

JA-18.1

24, 1.5/1.8mL microcentrifuge tubes

18,000 / 42,100

JA-20

24, 1.5/1.8mL microcentrifuge tubes

20,000 / 48,400

JA-30.50 Ti

24, 1.5mL microcentrifuge tubes

30,000 / 73,400

JS-4.0

104, 1.5mL microcentrifuge tubes

4, 96 deep-well plates

4,000 / 4,050

JS-4.3

104, 1.5/1.8mL microcentrifuge tubes

4, 96 deep-well plates

4,300 / 4,220

(3,250 / 1,924 with Micro Plus carriers)

JS-5.3

4, DNA kit 96-well filtration plates

5,300 / 6,130

JS-5.9

2, DNA kit 96-well filtration plates

5,900 / 6,570

JS-13.1

36 1.8mL microcentrifuge tubes

13,000 / 26,500

About Beckman Coulter

Beckman Coulter develops, manufactures and markets products that simplify, automate and innovate complex biomedical tests.

More than a quarter of a million Beckman Coulter instruments operate in laboratories around the world, supplying critical information for improving patient health and reducing the cost of care.


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Last updated: Mar 1, 2019 at 4:41 AM

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