On the Use of Buffy or Whole Blood for Obtaining DNA of High Quality and Functionality: What Is the Best Option?

Ferro, P.; Ortega-Pinazo, Jesús; Martínez, Beatriz; Jiménez, Álvaro; Gómez-Zumaquero, Juan M.; Hortas, Maria Luisa; Díaz, Tatiana

doi:10.1089/bio.2019.0024

Cited by 10 publications

(6 citation statements)

References 22 publications

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“…[16][17][18] Sample storage time has been reported to likely affect DNA concentration and purity due to nucleic acids natural degradation process. [19][20][21] Another DNA important variable is purity. We obtained a 260/280 ratio within a range from 1.64 to 2.05, with no statistically significant differences between the different groups that were analyzed.…”

Section: Discussionmentioning

confidence: 99%

Colony-forming units optimization and human papillomavirus detection in umbilical cord blood

Dimas-González¹,

Trejo-Gómora²,

Bonilla-Cisneros³

et al. 2023

GMM

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Introduction: Analysis of several parameters is required for adequate quality control in umbilical cord blood units (UCBU) when used for therapeutic purposes. Objective: To optimize colony-forming units (CFU) from clonogenic cultures and to detect the human papillomavirus (HPV) genome in UCBU. Methods: One hundred and forty-one umbilical cord blood (UCB), segment or CFU samples from UCBU clonogenic cultures were included. DNA extraction, quantification and endogenous GAPDH gene PCR amplification were carried out. Subsequently, HPV L1 gene was detected using the MY09/MY11 and GP5/GP6+ oligonucleotides. PCR products were analyzed with electrophoresis in agarose gel. CFU-extracted purified DNA was analyzed by electrophoresis in agarose gel, as well as some DNAs, using the sequence-specific priming technique. Results: CFU-extracted DNA concentration was higher in comparison with that of UCB (p = 0.0041) and that of the segment (p < 0.0001), as well as that of UCB in comparison with that of the segment (p < 0.0001). All samples were positive for GAPDH amplification and negative for MY09/MY/11 and GP5/GP6+. Conclusions: Cryopreserved UCBUs were HPV-negative. Obtaining CFU DNA from clonogenic cultures with high concentrations and purity is feasible.

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Section: Discussionmentioning

confidence: 99%

Colony-forming units optimization and human papillomavirus detection in umbilical cord blood

Dimas-González¹,

Trejo-Gómora²,

Bonilla-Cisneros³

et al. 2023

GMM

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“…В настоящем исследовании 4,5-летнее наблюдение также не выявило достоверного снижения концентрации ДНК при повторном измерении после хранения на -32 О С. Еще одним способом является хранение лейкоцитарного слоя (buffy coat) при -80 О C, полученного из образцов цельной крови, и дальнейшее выделение ДНК при необходимости. К недостаткам этого подхода можно отнести сложность автоматического процессинга лейкоцитарного слоя из образцов крови и меньший выход ДНК при длительном хранении по сравнению с образцами цельной крови, что может быть связано с положительным влиянием плазмы на консервацию ДНК [17].…”

Section: Discussionunclassified

Shelf life of whole blood samples in a biobank and the yield of deoxyribonucleic acid during genetic testing

Скирко¹,

Мешков²,

Ефимова³

et al. 2020

Cardiovasc Ther Prev

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Aim. To study the effect of the shelf life of frozen whole blood samples in a biobank on the amount of released deoxyribonucleic acid (DNA).Material and methods. The study included whole blood samples placed in tubes with the anticoagulant EDTA (ethylenediaminetetraacetic acid at a concentration of 1,8 mg/ml) from participants in the epidemiological study ESSE-RF-1 and ESSE-RF-2 and cohort studies conducted at the National Medical Research Center for Therapy and Preventive Medicine. The samples were stored in the biobank of the National Medical Research Center for Therapy and Preventive Medicine at temperature from -22О C to -32О C. The shelf life from blood collection to DNA extraction ranged from several weeks to 11 years. DNA was extracted using QIAamp DNA Blood Mini Kit (250) and 96 Blood Kit (Qiagen, Germany). Statistical analysis was performed using the R 3.6.1 software. To analyze the association of blood storage time with the logarithm of DNA concentration, a linear regression was used.Results. The analysis included data on the DNA concentration of 5405 samples. Multivariate regression showed that the blood shelf life was significantly associated with a decrease in concentration by 3,92% (3,16-4,68) for each year of storage (p <0,0001). For 509 samples, the DNA concentration was measured twice, immediately after isolation and after 4,5 years of DNA storage at -32О C. During storage, the concentration of DNA increased by an average of 2% (p=0,046).Conclusion. Long-term storage of whole blood samples at temperature from -22О C to -32О C is associated with a decrease in the DNA yield. Long-term storage of the isolated DNA at a temperature of -32О C is not associated with a decrease in its concentration.

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“…[16][17][18] Se ha reportado que el tiempo de almacenamiento de las muestras puede afectar la concentración y pureza del ADN debido al proceso de degradación natural de los ácidos nucleicos. [19][20][21] Otra variable importante del ADN es la pureza. Obtuvimos una relación 260/280 en un rango de 1.64 a 2.05, sin diferencias estadísticamente significativas entre los diferentes grupos analizados.…”

Section: B Aunclassified

Optimización de unidades formadoras de colonias y detección del virus de papiloma humano en sangre de cordón umbilical

Dimas-González

Trejo-Gómora

Bonilla-Cisneros

et al. 2021

GMM

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On the Use of Buffy or Whole Blood for Obtaining DNA of High Quality and Functionality: What Is the Best Option?

Cited by 10 publications

References 22 publications

Colony-forming units optimization and human papillomavirus detection in umbilical cord blood

Colony-forming units optimization and human papillomavirus detection in umbilical cord blood

Shelf life of whole blood samples in a biobank and the yield of deoxyribonucleic acid during genetic testing

Optimización de unidades formadoras de colonias y detección del virus de papiloma humano en sangre de cordón umbilical

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