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Microarray technology: the future of blood testing? Citations & References

  • Authors: Petrik J
  • Journal: Vox Sang
  • PubMed ID: 11339061

Comparison of detection of glucose-6-phosphate dehydrogenase deficiency using fluorescent spot test, enzyme assay and molecular method for prediction of severe neonatal hyperbilirubinaemia Citations & References

  • Authors: Wong, FL; Boo, NY; Ainoon, O; Wang, MK
  • Journal: SINGAPORE MEDICAL JOURNAL

Adenosine production by human B cells and B cell-mediated suppression of activated T cells. Citations & References

  • Authors: Saze Z, Schuler PJ, Hong CS, Cheng D, Jackson EK, Whiteside TL,
  • Journal: Blood
  • PubMed ID: 23678003

What sample types are compatible with QuantiGene Plex assays? Product FAQ

Answer

Whole blood, PaxGene blood, fresh tissue or FFPE tissues, cultured cells, and purified RNA have been tested with QuantiGene Plex assays. Other sample types including Tempus blood, bacteria, H&E stained sections, and blood spots are also compatible. Various sample prep protocols and kits are available.

Answer Id:: E19630

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Identification of low HBV-DNA levels by nucleic acid amplification test (NAT) in blood donors Citations & References

  • Authors: Dettori, S; Candido, A; Kondili, LA; Chionne, P; Taffon, S; Genovese, D; Iudicone, P; Miceli, M; Rapicetta, M
  • Journal: JOURNAL OF INFECTION

What is the recommended genomic DNA extraction methods used for the CytoScan assay? Product FAQ

Answer

Both blood and cell line sample sources have been tested with the CytoScan assay.
Methods that include boiling or strong denaturants are not acceptable because the DNA would be rendered single-stranded. Genomic DNA extracted using the following methods have been tested at ThermoFisher Scientific:

QIAGEN - Gentra Puregene Kit
5 PRIME - PerfectPure DNA Blood Kit
The CytoScan assay requires genomic DNA concentration >50 ng/µL. Therefore, the elution volumes for each of the kits will need to be adjusted accordingly to achieve the desired concentration.

Answer Id:: E13950

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Blood monitoring of Granzyme B and Perforin expression after intestinal transplantation: Considerations on clinical relevance Citations & References

  • Authors: Altimari, A; Gruppioni, E; Capizzi, E; Bagni, A; Corti, B; Fiorentino, M; Lazzarotto, T; Lauro, A; Pinna, AD; Ridolfi, L; Grigioni, WF; D'Errico-Grigioni, A
  • Journal: TRANSPLANTATION

Can the GeneChip Globin-Reduction kit be used for RNA isolated from starting materials different from whole blood? Product FAQ

Answer

The protocol has only been tested using RNA isolated using the PAXgene system. Theoretically, it might be beneficial to perform globin reduction if biological materials contaminated with blood are used for RNA isolation (e.g. aorta, heart). In determining whether this is necessary, we have found that globin levels corresponding to less than approximately 20% of the total poly (A)+ mRNA fraction do not require globin reduction prior to the GeneChip Target Labeling Assay. For more information, please refer to the technical note entitled "Globin Reduction Protocol: A Method for Processing Whole Blood RNA Samples for Improved Array Results" (http://media.affymetrix.com/support/technical/technotes/blood2_technote.pdf).

Answer Id:: E13484

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Evaluation of the conjunctival swab for canine visceral leishmaniasis diagnosis by PCR-hybridization in Minas Gerais State, Brazil. Citations & References

  • Authors: Ferreira Sde A; Ituassu LT; de Melo MN; de Andrade AS
  • Journal: Veterinary Parasitology
Catalog #

What is BVD virus (BVDV) in fetal bovine serum and how does it apply to cell culture applications? Product FAQ

Answer

BVD stands for Bovine Viral Diarrhea. It is one of the most common viral infections in cattle. It is estimated that 70 to 90 percent of the world's cattle population is seropositive for BVD. This virus can cause abnormalities and fetal abortions in cattle. Several strains of BVD exist, some of which are non-pathogenic.

Bovine serum is tested in accordance with 9 CFR, Section 113.53. The BVDV fluorescent antibody test is one of the required tests to meet Title 9 part 113.53 of the Code of Federal Regulations. However, the results of this test are somewhat subjective in the way they are scored. They are scored 0 to +4, based on the level of observed fluorescence. Most samples will have a detectable level of BVDV antigen due to its prevalence in the bovine population. The FA part of the testing can reveal the presence of non-cytopathic BVD strains. The Cytopathogenic and Hemadsorbing Agents testing is used to determine the release of bovine serum, regardless of the BVDV result. We currently report the BVDV results as “Tested”. Any live cytopathic bovine viruses (including the cytopathic BVDV strain) would be revealed in the testing for cytopathogenic agents. In this portion of the testing, the cultures are microscopically monitored for evidence of inclusion bodies, abnormal number of giant cells, or other cytopathology indicative of cell abnormalities. The hemadsorption assay detects the presence of hemagglutinating viruses. The viral hemagglutinin would induce clumping of red blood cells. If a positive result in the cytopathogenic agents or hemadsorbing agents assays is reported, the material would be failed and therefore would not be released by Quality Assurance.

Answer Id:: E11874

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Habitual Energy Expenditure Modifies the Association Between NOS3 Gene Polymorphisms and Blood Pressure. Citations & References

  • Authors: Vimaleswaran KS; Franks PW; Barroso I; Brage S; Ekelund U; Wareham NJ; Loos RJ
  • Journal: American Journal of Hypertension : Journal of the American Society of Hypertension

Development of a blood based breast cancer test for Indian population Citations & References

  • Authors: Tobin, D; Baardsen, K; Lindahl, T; Kauczynska, M; Punia, DP; Kumar, Y; Desai, C; Shroff, C; Borresen-Dale, AL; Sharma, P
  • Journal: CANCER RESEARCH

How do you prepare blood cells for chromosome analysis? Product FAQ

Answer

Phytohemagglutinin Assay:

Lymphocytes are differentiated cells which normally do not undergo subsequent cell divisions. By culturing lymphocytes in the presence of a mitogen (KaryoMAX Phytohemagglutinin (M-Form) (PHA), Cat. No. 10576), they are stimulated to replicate their DNA and enter into mitosis. After an optimum time of the cells being cultured (46 h for a newborn and 68 h for an adult), a mitotic inhibitor, KaryoMAX COLCEMID Solution (Cat. No. 15210 or 15212), is added to the lymphocyte culture for 20 min. The addition of COLCEMID to dividing cells acts to prevent the synthesis of spindle fibers and, therefore, to stop mitosis in metaphase. Metaphase is the optimum phase of mitosis for microscopically visualizing the chromosomes. By submitting cells to a hypotonic solution and a series of fixation steps, metaphase chromosomes can be microscopically observed and analyzed.

As a quality control measure, each lot of Phytohemagglutinin is tested as a chromosome reagent for the examination of metaphase spreads used for cytogenetic studies. This reagent is evaluated by supplementation to an approved, non-phytohemagglutinin containing chromosome medium. These samples are then supplemented with freshly collected human peripheral blood and have been found to be acceptable in their ability to produce blastogenesis with human lymphocytes when compared to a previously tested control.

Test Procedure:

1. The required volume of peripheral blood is collected aseptically in a sodium heparinized vacutainer tube or syringe.
2. Add 10 ml of either PB-MAX Karyotyping Medium (Cat. No. 10386) to each sterile T-25 flask to be set up for the assay.
3. Add 0.75ml blood to each tube.
4. Incubate flask in CO2 incubator for 48 to 68 h with caps loose.
5. Add 0.05-0.1 ug/ml COLCEMID to each flask for a 15 minute incubation.
6. After 15 min, transfer flask contents to a 15 ml centrifuge tube and spin down at 1,200 rpm for 5 min.
7. Remove supernatant and resuspend pellet.
8. Add 10 ml of 0.068 M KCl to pellet and gently mix. Allow to sit at room temperature for 15 min. Add 0.5 ml of fixative (three parts absolute methanol to one part glacial acetic acid). Gently mix with pipette.
9. Centrifuge for 5 min at 1,200 rpm. Aspirate off supernatant. Add 10 ml of fixative, mix, and let sit at room temperature for 10 min.
10. Repeat centrifugation step. Add 5 ml of fixative, mix, and let sit for 10 min at room temperature.
11. Centrifuge and aspirate off supernatant. Add 5 ml of fixative and incubate for 10 min at 4°C.
12. Centrifuge at 1,200 rpm for 7 min. Aspirate off supernatant. Gently resuspend in fixative.
13. Prepare slide by placing 6 to 8 drops of cell suspension on slide. Use warming plate to dry slides.
14. Once all traces of moisture have disappeared, you can begin staining procedure:
- Stain with Giemsa Stain for 6 to 8 min, or any other appropriate stain.
- Remove slides from stain and rinse under running distilled water.
- Allow slides to thoroughly dry and examine slides for well-spread metaphases.

Record number of cells and at the same time record the number of these cells which are in metaphase. At least 500 cells must be counted for a valid assay. Statistically determine the Mean Mitotic Index (a quantitative measure) and Mean Banding Resolution (a qualitative measure). The test sample values must favorably compare to the reference control values.

Answer Id:: E4306

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Does the sample input for the TrueMark MSI Assay need to be FFPE samples or can I use blood (cfDNA)? Product FAQ

Answer

The TrueMark MSI Assay has only been tested with FFPE samples at this time.

Answer Id:: E18145

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What endogenous control to use for human miRNA from blood (serum, plasma)? Product FAQ

Answer

Small RNAs such as snRNAs or snoRNAs are usually not present in serum or other body fluids. Spike in controls can be used to monitor sample preparation. Any miRNA that is present in your serum samples can be used as a control as long as it is stably expressed across all the sample types in your study. You can refer to the literature for candidate miRNAs to test or you can select of a control from your data set to use in your analysis.

Answer Id:: E7052

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