A comparative study of the quality of DNA obtained from fresh frozen and formalin-fixed decalcified paraffin-embedded bone marrow trephine biopsy specimens using two different methods

Talaulikar, Dipti; Gray, Jennifer; Shadbolt, Bruce; McNiven, Michelle; Dahlstrom, Jane E.

doi:10.1136/jcp.2006.045294

Cited by 30 publications

(23 citation statements)

References 17 publications

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“…Table 2) were amplified with even better success and also sequenced. Recent studies revealed lowest percentage of amplification for fragments around 600 bp: in thirteen samples tested Talaulikar et al (2008) achieved 10% of amplification and Gillio-Tos et al (2007) were able to amplify 5% of 365 gDNA samples tested.…”

Section: Discussionmentioning

confidence: 95%

An efficient protocol for genomic DNA extraction from formalin-fixed paraffin-embedded tissues

Santos

Sá

Bastos

et al. 2009

Research in Veterinary Science

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Formalin-fixed paraffin-embedded tissues (FFPET) represent the largest source of archival biological material available for genomic studies. In this work we present an advanced protocol for extraction of high quality DNA from FFPET that can be applied in several molecular studies. Although cat mammary tumours (CMT) are the third most frequent tumour in cats the recovery of significant number of samples for molecular studies are in some way restricted to FFPET samples. We were able to obtain high quality DNA from FFPET of thirty six CMT that were subjected to prefixation and fixation processes routinely used in the veterinary hospitals. The quality of DNA obtained was tested by PCR amplification using six sets of primers that amplify single-copy fragments. The DNA fragments obtained were further sequenced. This protocol was able to provide FFPET gDNA that can be amplified and sequenced for larger fragments up to 1182 bp.

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Section: Discussionmentioning

confidence: 95%

An efficient protocol for genomic DNA extraction from formalin-fixed paraffin-embedded tissues

Santos

Sá

Bastos

et al. 2009

Research in Veterinary Science

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“…In our study, however, DNA was extracted from freshly frozen tissue from which fresh biopsies are better digested and yield good quality DNA [14], and the amplified product was detected by ELISA as an established quantitative and qualitative method (PCR detection limit was 20 DNA copies/reaction).…”

Section: Discussionmentioning

confidence: 99%

Are human herpes viruses associated with autoimmune thyroid disease?

AL-Zarzour

Monem

2011

J Infect Dev Ctries

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Introduction: Autoimmune diseases are complex diseases with genetic, endogenous and environmental etiologies. Viral infections have been postulated as one of the factors that may be the trigger of autoimmune diseases. Methodology: Thyroid peroxidase (TPO) and thyroglobulin (TG) antibodies were measured before thyroidectomy in 100 subjects by chemiluminescence method, 50 of whom were autoimmune thyroid diseases (AITD) patients and 50 of whom were multinodular goiter (MNG) patients used as a control group. Fresh thyroid samples were collected from all 100 subjects after thyroidectomy to investigate the DNAs of herpes simplex viruses types 1 and 2 (HSV-1, HSV-2), Varicella Zoster virus (VZV), Epstein-Barr virus (EBV), Cytomegalovirus (CMV) and human herpes virus type 6 (HHV-6) by PCR. Results: The DNA of HSV-1, HSV-2, VZV, EBV, CMV and HHV-6 were detected in neither the patient group nor in the control group. The mean values of anti-TPO and anti-TG antibodies ranged within 9.5-2000 units/ml (527.8 ± 617.4) and 108-5000 units/ml (1458.2 ± 1774.1) in the AITD patients group, respectively. A statistically significant difference of the mean level of anti-TPO and anti-TG antibodies among the two groups was found (p value < 0.05). Conclusions: The possible role of human herpes viruses in the pathogenesis of AITD is not supported by our study; hence our raised question stays open for more investigation on more patients and in different parts of the country using different sizes and sites of biopsies.

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“…Some of these limited samples, however, fail molecular testing mostly due to scant cellularity. In addition, molecular analysis of bone metastasis is difficult due to the decalcification process that negatively influences the quality of DNA [17,18]. Recently two monoclonal antibodies, specific to the two most common forms of mutated EGFR protein have become commercially available [19].…”

Section: Introductionmentioning

confidence: 99%

Use of mutation specific antibodies to detect EGFR status in small biopsy and cytology specimens of lung adenocarcinoma

et al. 2012

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A comparative study of the quality of DNA obtained from fresh frozen and formalin-fixed decalcified paraffin-embedded bone marrow trephine biopsy specimens using two different methods

Abstract: Although amplifiable DNA can be extracted from both fresh-frozen and FFDPE trephine samples for IgH/IgL analysis, freshly frozen specimens are superior as a source of template DNA, especially for higher base pair PCR products.

Cited by 30 publications

References 17 publications

An efficient protocol for genomic DNA extraction from formalin-fixed paraffin-embedded tissues

An efficient protocol for genomic DNA extraction from formalin-fixed paraffin-embedded tissues

Are human herpes viruses associated with autoimmune thyroid disease?

Use of mutation specific antibodies to detect EGFR status in small biopsy and cytology specimens of lung adenocarcinoma

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