Comparison of three different kits for extraction of high-quality RNA from frozen blood.

Author(s): Kim JH, Jin HO, Park JA, Chang YH, Hong YJ, Lee JK

Publication: Springerplus, 2014, Vol. 3, Page 76

PubMed ID: 24567882 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 RNA extraction method from blood specimens on yield, purity, RIN, and Ct values. 15 blood specimens were collected into EDTA tubes and stored at -80 degrees C for 5 years before extraction and analysis.

   Summary of Findings:

   The highest average RNA yields from frozen blood were obtained using TRI reagent (15.6 ug/mL) followed by NucleoSpin (9.0 ug/mL) and lastly PAXgene (3.1 ug/mL). RNA extraction with TRI reagent had the lowest purity (1.7), as measured by A260/A280, with PAXgene and NucleoSpin-extracted RNA giving similarly higher levels of purity (2.0). Lower A260/A230 results were obtained for PAXgene and TRI reagent-extracted RNA than for NucleoSpin-extracted RNA. RNA extracted with Tri reagent gave an average RIN score of 3.2 while that extracted by PAXgene or NucleoSpin had higher RIN scores (6.0 and 6.4, respectively). Ct values were generally highest for RNA extracted with TRI reagent and lowest for RNA extracted with NucleoSpin.

   Biospecimens

   • Bodily Fluid - Blood

   Preservative Types

   • Frozen

   Diagnoses:

   • Not specified

   Platform:

   Analyte	Technology Platform
   RNA	Automated electrophoresis/Bioanalyzer
   RNA	Spectrophotometry
   RNA	Real-time qRT-PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Analyte Extraction and Purification	Analyte isolation method	TRI reagent PAXgene NucleoSpin
   Real-time qRT-PCR Specific	Targeted nucleic acid	18s rRNA Beta actin RPLP0 HPRT1

   View more