GENOMIC DNA PREP KIT for BLOOD

ORDERING INFORMATION

Catalog #	Product	Amount	Price (EUR)
# GEN 50 B	Genomic DNA Prep Kit for Blood	50 preparations
# GEN 250 B	Genomic DNA Prep Kit for Blood	250 preparations

PRODUCT COMPONENTS

Buffer	# GEN 50 B	# GEN 250 B
Blood Lysis Buffer	50 ml	250 ml
Binding Buffer	35 ml	175 ml
Column Activation Solution	6 ml	30 ml
Elution Buffer	5 ml	25 ml
Spin Columns & 2ml Tubes	50	250

DESCRIPTION

The spin column based Blood DNA Preparation Kit is designed for rapid and high purity isolation of genomic DNA from whole blood. The spin column based method completely removes PCR inhibitors such as divalent cations and proteins resulting in a high purity preparation of genomic DNA. There is no use of phenol or chloroform, handling is safe and does not produce any harmful waste. Column based genomic DNA purification kits yield up to 30 μg DNA sized from 200 bp to 50 kb per preparation.

FEATURES

• Spin column type
• Purification size: 200 bp - 50 kb
• DNA amounts up to 30 µg

APPLICATIONS

• real-time PCR
  
• PCR
• southern blot analysis
  
• genotyping
• discovery or validation of SNP/SSR marker
  

BASIC PROTOCOL

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

• for 50 preparations (# GEN 50 B): 60 ml
• for 250 preparations (# GEN 250 B): 300 ml

1. Cell Lysis

• Mix 200 µl (1 volume) of whole human blood with 1 ml (5 volumes) of Blood Lysis Buffer in an appropriately sized tube (not provided).
• Incubate for 10 min on ice.
• Mix by vortexing briefly 2-3 times during incubation.
• Centrifuge at 12,000 g for 10 min at RT.
• Remove or discard supernatant completely.

2. Adding Binding Buffer

• Add 600 µl Binding Buffer to pellet.
• Vortex briefly.

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

• Pipette the lysate directly into the Spin Column.
• Centrifuge for 1 min at 12,000 g.

5. Primary Washing

• Add 500 µl Washing Buffer (80% Ethanol) into the Spin Column.
• Centrifuge for 30 sec at 12,000 g.
• Discard the flow-through.

6. Secondary Washing

• Add 500 µl Washing Buffer into the Spin Column.
• Centrifuge for 30 sec at 12,000 g.
• Discard the flow-through.
• Centrifuge again at 12,000 g for 1 min to remove residual Washing Buffer.
  
• Discard the 2 ml wash tube and place the column in the elution tube.
  

7. Elution of DNA

• Add 40-50 μl Elution Buffer to the center of the column.
• Incubate at room temperature for 1 min.
• Centrifuge at 12,000 g for 2 min to elute DNA.
• Store DNA at 4°C (for prolonged storage, keep it at -20°C or -80°C).

STORAGE

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