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GEL PURIFICATION SYSTEM - Column Type

ORDERING INFORMATION

Catalog #

Product

Amount

Price (EUR)

# GEL 50
ROVALAB Gel Purification System
50 preparations

43,50

# GEL 250
ROVALAB Gel Purification System

250 preparations

176,50

PRODUCT COMPONENTS

Buffer

# GEL 50

# GEL 250

Binding Buffer

75 ml

2 x 185 ml

Column Activation Solution

6 ml

30 ml

Elution Buffer

6 ml

30 ml

Spin Columns & 2ml Tubes

50

250

DESCRIPTION

ROVALAB Gel Purification System is designed for high-yield recovery of DNA from agarose gel with simultaneous removal of primer-dimers, primers, nucleotides, proteins, salt, agarose, ethidium bromide and other impurities. The preparation is based on a silicamembrane technology for binding DNA in high-salt and elution in low-salt buffer. The kit provides a simple and efficient way to purify DNA in a size range between 100 bp and 10 kb. It requires no organic extractions or precipitation and guarantees high and stable recovery rates.

FEATURES

  • High Yield: > 80%
  • Spin column type
  • Purification size: 100 bp - 10 kb
  • DNA amounts up to 20 g
  • Sequencing grade

APPLICATIONS

  • General sequencing
  • PCR
  • Cloning
  • Restriction enzyme digestion

BASIC PROTOCOL

The agarose gel is dissolved in the chaotropic Extraction Buffer followed by a simple binding, washing and eluting procedure.

Before start, prepare an 80% Ethanol (V/V) solution as Washing buffer:

  • for 50 preparations (# GEL 50): 80 ml
  • for 250 preparations (# GEL 250): 400 ml
The additional use of Isopropanol (not included) is recommended for fragments smaller than 200 bp or larger than 5 kbp.The optional secondary washing step minimizes the salt content of the purification product but may significantly reduce the yield of DNA fragments <200 bp.

1. Excision of the Gel

  • Cut the area of gel containing the DNA fragment.
  • Transfer the excised gel to a clean 1.5 ml microtube.

2. Sample Preparation

  • Add 3 volumes of Extraction Buffer to 1 volume of the sliced gel. For example, add 300 l Extraction Buffer to each 100 mg (approx. 100 μl) gel. For gels containing >2.5% agarose, add 6 volumes of Extraction Buffer per gel volume.
  • Incubate at 60C for 10 min with occasional mixing to ensure gel dissolution.
  • For DNA fragment sizes smaller than 200 bp or larger than 5 kbp and to enhance yield, add 1 volume Isopropanol per gel volume to the dissolved gel and mix well.

3. Column Activation

  • Place a Spin Column into a 2 ml collection tube.
  • Add 100 l of Column Activation Solution into the Spin Column.
  • Centrifuge at 10,000 g for 30 sec in a microcentrifuge.

4. Column Loading

  • Apply the sample mixture from step 2 into the activated Spin Column.
  • Centrifuge at 10,000 g for 30 sec in a microcentrifuge.
  • Discard the flow-through.

5. Column Washing

  • Place the DNA loaded Spin Column into the used 2 ml tube.
  • Apply 700 l of Washing Buffer (80% Ethanol) to the Spin Column.
  • Centrifuge at 10,000 g for 30 sec and discard the flow-through.
  • Optional Secondary Washing: Recommended only for DNA >200 bp, if highly purified DNA (for DNA sequencing, transfection etc.) is required.
    • Add 700 l of Washing Buffer to the Spin Column.
    • Centrifuge at 10,000 g for 30 sec and discard the flow-through.
  • Centrifuge again for 2 min to remove residual Washing Buffer.

6. Elution

  • Place the Spin Column into a clean 1.5 ml microtube (not provided in the kit).
  • Add 30-50 μl Elution Buffer to the center of the column membrane.
  • Incubate at room temperature for 1 min.
  • Centrifuge at 10,000 g for 1 min to elute DNA.

STORAGE

Store at room temperature (15 - 25 C) for 12 months.