Evaluation of Maxwell(®) 16 for automated DNA extraction from whole blood and formalin-fixed paraffin embedded (FFPE) tissue.

Author(s): Khokhar SK, Mitui M, Leos NK, Rogers BB, Park JY

Publication: Clin Chem Lab Med, 2012, Vol. 50, Page 267-272

PubMed ID: 22022984 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the effects of DNA isolation method on yield and amplificability of DNA from EDTA-blood.

   Summary of Findings:

   A significantly higher average DNA yield was obtained from blood extracted with the Maxwell 16 LEV blood DNA kit (118.68 ng/ul) than with the NucliSens EasyMag Blood DNA kit (21.33 ng/ul, p<0.0001) or the QIAamp DNA Blood mini kit (45.69 ng/ul, p<0.0001). The A260/280 ratio was 1.84 for specimens extracted with Maxwell 16 LEV blood DNA kit and the QIAamp DNA Blood mini kit, but it was only 1.58 when DNA was extracted using NucliSens EasyMag Blood DNA kit. Beta-actin was successfully amplified from all 10 blood specimens, regardless of extraction method.

   Biospecimens

   • Bodily Fluid - Blood

   Preservative Types

   • None (Fresh)

   Diagnoses:

   • Not specified

   Platform:

   Analyte	Technology Platform
   DNA	PCR
   DNA	Spectrophotometry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Analyte Extraction and Purification	Analyte isolation method	QIAamp DNA Blood mini kit Maxwell 16 LEV blood DNA kit NucliSens EasyMag Blood DNA kit
   PCR Specific	Targeted nucleic acid	Beta-actin

   View more

2. Study Purpose

   The purpose of this study was to determine the effects of DNA isolation method and amplicon size on PCR success using FFPE liver specimens. DNA was extracted from 20 micron sections.

   Summary of Findings:

   Significantly higher DNA yields were obtained from FFPE sections extracted using the Qiagen AllPrep FFPE DNA/RNA kit (129 ng/ul) than the Qiagen AllPrep DNA/RNA kit (7.9 ng/ul, p<0.0001) or the Maxwell 16 FFPE tissue LEV DNA kit (17.42, p<0.0001). The A260/280 ratio was 1.9 or greater for DNA extracted from FFPE liver sections using each of the three DNA extraction kits. A 286 bp fragment of CDKL5 exon 3 was successfully amplified using DNA from all 10 FFPE liver sections, regardless of extraction method, but a while 402 bp fragment of exon 16 of CDKL5 was successfully amplified in all 10 specimens extracted with the Maxwell 16 FFPE tissue LEV DNA kit, it was not amplified in all 10 specimens extracted by the other methods. In contrast, a 618 bp fragment of exon 5 was amplified in 2 of 10 FFPE sections extracted with Qiagen AllPrep FFPE DNA/RNA kit, but the same fragment was not amplified in DNA from FFPE sections extracted with Maxwell 16 FFPE tissue LEV DNA kit or the Qiagen AllPrep DNA/RNA kit.

   Biospecimens

   • Tissue - Liver

   Preservative Types

   • Formalin

   Diagnoses:

   • Not specified

   Platform:

   Analyte	Technology Platform
   DNA	PCR
   DNA	Spectrophotometry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Analyte Extraction and Purification	Analyte isolation method	Qiagen AllPrep DNA/RNA kit Qiagen AllPrep FFPE DNA/RNA kit Maxwell 16 FFPE tissue LEV DNA kit
   PCR Specific	Targeted nucleic acid	CDKL5 exon 3 CDKL5 exon 5 CDKL5 exon 16
   PCR Specific	Length of gene fragment	286 bp 402 bp 618 bp

   View more