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Rapid DNA extraction from dried blood spots on filter paper: potential applications in biobanking.

Choi EH, Lee SK, Ihm C, Sohn YH - Osong Public Health Res Perspect (2014)

Bottom Line: DNA was stable in DBSs that were stored at room temperature or frozen.The housekeeping genes GAPDH and beta-actin were used as positive standards for polymerase chain reaction (PCR) validation of general diagnostic screening.Our simple and convenient DBS storage and extraction methods are suitable for diagnostic screening by using very small volumes of blood collected on filter paper, and can be used in biobanks for blood sample storage.

View Article: PubMed Central - PubMed

Affiliation: Eulji Medi-Bio Research Institute (EMBRI), Daejeon, Korea.

ABSTRACT

Objectives: Dried blood spot (DBS) technology is a microsampling alternative to traditional plasma or serum sampling for pharmaco- or toxicokinetic evaluation. DBS technology has been applied to diagnostic screening in drug discovery, nonclinical, and clinical settings. We have developed an improved elution protocol involving boiling of blood spots dried on Whatman filter paper.

Methods: The purpose of this study was to compare the quality, purity, and quantity of DNA isolated from frozen blood samples and DBSs. We optimized a method for extraction and estimation of DNA from blood spots dried on filter paper (3-mm FTA card). A single DBS containing 40 μL blood was used.

Results: DNA was efficiently extracted in phosphate-buffered saline (PBS) or Tris-EDTA (TE) buffer by incubation at 37°C overnight. DNA was stable in DBSs that were stored at room temperature or frozen. The housekeeping genes GAPDH and beta-actin were used as positive standards for polymerase chain reaction (PCR) validation of general diagnostic screening.

Conclusion: Our simple and convenient DBS storage and extraction methods are suitable for diagnostic screening by using very small volumes of blood collected on filter paper, and can be used in biobanks for blood sample storage.

No MeSH data available.


Related in: MedlinePlus

Yield of ssDNA from dried blood spots stored at −80°C and room temperature. ssDNA = single-stranded DNA.
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fig5: Yield of ssDNA from dried blood spots stored at −80°C and room temperature. ssDNA = single-stranded DNA.

Mentions: As the filter paper storage method provides higher ssDNA yields than the frozen-liquid method at room temperature, we investigated how the storage temperature affected yields. As shown in Figure 5, ssDNA yields were compared between filter paper storage temperatures and times; there was little difference between the amounts of ssDNA extracted. However, there was a difference of 8% in the ssDNA yields between 2- and 4-week storage, while there was a 1% difference in ssDNA yields from samples stored at room temperature for the same period. Therefore, storage at room temperature was best for samples on filter paper.


Rapid DNA extraction from dried blood spots on filter paper: potential applications in biobanking.

Choi EH, Lee SK, Ihm C, Sohn YH - Osong Public Health Res Perspect (2014)

Yield of ssDNA from dried blood spots stored at −80°C and room temperature. ssDNA = single-stranded DNA.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4281615&req=5

fig5: Yield of ssDNA from dried blood spots stored at −80°C and room temperature. ssDNA = single-stranded DNA.
Mentions: As the filter paper storage method provides higher ssDNA yields than the frozen-liquid method at room temperature, we investigated how the storage temperature affected yields. As shown in Figure 5, ssDNA yields were compared between filter paper storage temperatures and times; there was little difference between the amounts of ssDNA extracted. However, there was a difference of 8% in the ssDNA yields between 2- and 4-week storage, while there was a 1% difference in ssDNA yields from samples stored at room temperature for the same period. Therefore, storage at room temperature was best for samples on filter paper.

Bottom Line: DNA was stable in DBSs that were stored at room temperature or frozen.The housekeeping genes GAPDH and beta-actin were used as positive standards for polymerase chain reaction (PCR) validation of general diagnostic screening.Our simple and convenient DBS storage and extraction methods are suitable for diagnostic screening by using very small volumes of blood collected on filter paper, and can be used in biobanks for blood sample storage.

View Article: PubMed Central - PubMed

Affiliation: Eulji Medi-Bio Research Institute (EMBRI), Daejeon, Korea.

ABSTRACT

Objectives: Dried blood spot (DBS) technology is a microsampling alternative to traditional plasma or serum sampling for pharmaco- or toxicokinetic evaluation. DBS technology has been applied to diagnostic screening in drug discovery, nonclinical, and clinical settings. We have developed an improved elution protocol involving boiling of blood spots dried on Whatman filter paper.

Methods: The purpose of this study was to compare the quality, purity, and quantity of DNA isolated from frozen blood samples and DBSs. We optimized a method for extraction and estimation of DNA from blood spots dried on filter paper (3-mm FTA card). A single DBS containing 40 μL blood was used.

Results: DNA was efficiently extracted in phosphate-buffered saline (PBS) or Tris-EDTA (TE) buffer by incubation at 37°C overnight. DNA was stable in DBSs that were stored at room temperature or frozen. The housekeeping genes GAPDH and beta-actin were used as positive standards for polymerase chain reaction (PCR) validation of general diagnostic screening.

Conclusion: Our simple and convenient DBS storage and extraction methods are suitable for diagnostic screening by using very small volumes of blood collected on filter paper, and can be used in biobanks for blood sample storage.

No MeSH data available.


Related in: MedlinePlus