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 µg
• 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:

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 µl 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 60°C 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 µl 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 µl 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 µl 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.