Independent, peer-reviewed papers highlight the usefulness of FTA™ cards in specific DNA applications

We have identified a series of independent, peer-reviewed papers that highlight the usefulness of FTA™ cards in specific DNA applications, and in which you will find useful ideas, hints and tips for the use of FTA cards in your sample collection. The abstracts and conclusions from the first two papers are included below.

1. Performance of Identifiler® Direct and PowerPlex® 16 HS on the Applied Biosystems 3730 DNA Analyzer for processing biological samples archived on FTA™ cards
Laurin et al, Volume 6, Issue 5, September 2012, Pages 621–629 

Abstract

FTA™ card with e-Core™ sampling 

Direct amplification of STR loci from biological samples collected on FTA™ cards without prior DNA purification was evaluated using Identifiler® Direct and PowerPlex 16 HS in conjunction with the use of a high throughput Applied Biosystems 3730 DNA Analyzer. In order to reduce the overall sample processing cost, reduced PCR volumes combined with various FTA™ disk sizes were tested. Optimized STR profiles were obtained using a 0.53 mm disk size in 10 µl PCR volume for both STR systems. These protocols proved effective in generating high quality profiles on the 3730 DNA Analyzer from both blood and buccal FTA™ samples. Reproducibility, concordance, robustness, sample stability and profile quality were assessed using a collection of blood and buccal samples on FTA™ cards from volunteer donors as well as from convicted offenders. The new developed protocols offer enhanced throughput capability and cost effectiveness without compromising the robustness and quality of the STR profiles obtained. These results support the use of these protocols for processing convicted offender samples submitted to the National DNA Data Bank of Canada. Similar protocols could be applied to the processing of casework reference samples or in paternity or family relationship testing.

• ABI & Promega have configured robust PCR mixes to overcome inhibitors when FTA™ punches are added directly to PCR without punch washing
• No-wash workflows increase convenience and sample throughput when combined with optimised chemistries
• These premium products have delivered improved full-profile first-pass rates in forensic DNA reference laboratories
• By decreasing punch sizes below 1.2 mm diameter, labs are moving from 25 µl towards 10 µl STR miniaturised reactions
• Miniaturisation of reaction mixes has been sought to drive down costs

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2. The use of FTA™ cards for preserving unfixed cytological material for high-throughput molecular analysis.  Saleg M. A et al. Cancer cytopathology Volume 120, Issue 3, 25 June 2012, Pages 206-214

Abstract
BACKGROUND: Novel high-throughput molecular technologies have made the collection and storage of cells and small tissue specimens a critical issue. The FTA™ card provides an alternative to cryopreservation for biobanking fresh unfixed cells. The current study compared the quality and integrity of the DNA obtained from 2 types of FTA™ cards (Classic and Elute) using 2 different extraction protocols (‘‘Classic’’ and ‘‘Elute’’) and assessed the feasibility of performing multiplex mutational screening using fine-needle aspiration (FNA) biopsy samples. METHODS: Residual material from 42 FNA biopsies was collected in the cards (21 Classic and 21 Elute cards). DNA was extracted using the Classic protocol for Classic cards and both protocols for Elute cards. Polymerase chain reaction for p53 (1.5 kilobase) and CARD11 (500 base pair) was performed to assess DNA integrity. RESULTS: Successful p53 amplification was achieved in 95.2% of the samples from the Classic cards and in 80.9% of the samples from the Elute cards using the Classic protocol and 28.5% using the Elute protocol (P = .001). All samples (both cards) could be amplified for CARD11. There was no significant difference in the DNA concentration or 260/280 purity ratio when the 2 types of cards were compared. Five samples were also successfully analysed by multiplex MassARRAY® spectrometry, with a mutation in KRAS found in 1 case. CONCLUSIONS: High molecular weight DNA was extracted from the cards in sufficient amounts and quality to perform high-throughput multiplex mutation assays. The results of the current study also suggest that FTA™ Classic cards preserve better DNA integrity for molecular applications compared with the FTA™ Elute cards.

• Typical preservation of biopsy tissues with FFPE have been reported to cause cross-links and damage DNA
• The Elute protocol with water (98°C; 30min) produces ssDNA and can lower DNA quality with additional fragmentation
• Larger amplicons from higher quality DNA is achieved with FTA™ compared to FTA™ Elute

Download the first chapter of our eBook, on sample collection, shipping and storage with FTA cards.

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The Polymerase Chain Reaction (PCR) is covered by patents owned by Roche Molecular Systems and F Hoffmann-La Roche Ltd. All third party trademarks are the property of their respective owners. Identifiler is a registered trademark of Applied Biosystems. PowerPlex is a registered trademark of Promega Corporation. FTA is a trademark of GE Healthcare companies.
Regulatory Note: The study/data quoted in the paper above were not GEHC studies and GEHC makes no claims on the FTA or FTA Elute instructions for use regarding any down-stream testing of the sample. The down-stream testing and any validation of these tests is the responsibility of the end-user