Precautions To Avoid Contamination; Samples Collection And Transport; Rna Extraction - INGENASA INgene q IBV SYBR Instrucciones De Uso

5. PRECAUTIONS TO AVOID CONTAMINATION

The following points should be read with care:
Disposable items must be RNAse and DNAse-free.
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Use RNAse and DNAse-free distilled, autoclaved water (25 min., 120ºC).
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Use sterile filtered tips.
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Ensure good homogenisation of the kit components once defrosted.
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Maintain the A and B mixes stored in ice at all times.
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Repeated cycles of freezing and defrosting may reduce the sensitivity of the reagents. Protect them from exposure
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to light until use.
To avoid contaminations giving rise to false positives it is important to:
Physically separate the positive PCR control from the remaining reagents of the kit.
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To carry out any handling of samples to be tested in a different location/room from the one where the amplified

products are being analyzed.
Add the positive PCR control in a different location/room from the one where the mix is added and where the
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samples to be tested are being handled.

6. SAMPLES COLLECTION AND TRANSPORT

INgene q IBV SYBR is suited for the detection (optionally quantification) of RNA of Infectious Bronchitis virus (IBV) in
biological samples.
The assay has been standardised and contrasted with cecal and tracheal swab, trachea and kidney samples.
Samples must be cooled from the moment they are obtained until they are processed. Processing needs to occur within 24
hours after obtaining the samples. In case a longer time is envisaged, we recommend freezing the samples.

7. RNA EXTRACTION

Any extraction procedure should be used which yields good-quality of the extracted material.
In case you have no optimized extraction procedures, INGENASA could provide extraction protocol and buffer under
request.
Ingenasa has tested the following methods for the RNA extraction:
Method of Chomczynski and Sacchi (Phenol extraction).
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Automatic magnetic methods (Mag MAX
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Column purification methods: QIAamp® Viral RNA Mini Kit (QIAGEN), High Pure Viral Nucleic Acid kit (ROCHE).
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There are several commercial houses that provide high quality extraction kits that could be used following the
manufacturer´s instructions, such as: Life-technologies, ROCHE, Quiagen, Marchery Nalgene, others.
CAUTION: If amplification is not going to be done immediately, preserve the RNA at -20° C.
INSTRUCTIONS – ENGLISH
TM
Total NA isolation kit, Ref AM1840).
(Versión 1410-14) – PAG.9.