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Protocol for the Use of Polymerase Chain Reaction in the Detection of Intraocular Large B-Cell Lymphoma in Ocular Samples  Aires Lobo, Narciss Okhravi,

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Presentation on theme: "Protocol for the Use of Polymerase Chain Reaction in the Detection of Intraocular Large B-Cell Lymphoma in Ocular Samples  Aires Lobo, Narciss Okhravi,"— Presentation transcript:

1 Protocol for the Use of Polymerase Chain Reaction in the Detection of Intraocular Large B-Cell Lymphoma in Ocular Samples  Aires Lobo, Narciss Okhravi, Peter Adamson, Brian J Clark, Susan Lightman  The Journal of Molecular Diagnostics  Volume 9, Issue 1, Pages (February 2007) DOI: /jmoldx Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

2 Figure 1 Amplification of Fr2, Fr3, and Bcl-2 t(14;18) PCR product from purified genomic DNA. A: Varying quantities of DOHH B-cell lymphoma genomic DNA were subjected to PCR amplification using Fr2 primers, which detected 10 pg of B-cell lymphoma DNA. PCR products (10 μl) were resolved on 8% PAGE/TBE gels and visualized after SYBR Gold staining under UV illumination. B: Seminested amplification of Fr2 PCR product generated from purified genomic DNA detected 10 fg of B-cell DNA, using seminested Fr2 primers. PCR products (10 μl) were resolved on 8% PAGE/TBE gels and visualized after ethidium bromide staining under UV illumination. C: Varying quantities of RAJI B-cell lymphoma genomic DNA were subjected to PCR amplification using Fr3 primers, which detected 100 pg of B-cell lymphoma DNA. PCR products (10 μl) were resolved on 8% PAGE/TBE gels and visualized after SYBR Gold staining under UV illumination. D: Seminested amplification of Fr3 PCR product generated from purified genomic DNA detected 10 pg of B-cell DNA, using seminested primers Fr3 primers. PCR products (10 μl) were resolved on 8% PAGE/TBE gels and visualized after ethidium bromide staining under UV illumination. E: Varying quantities of DOHH B-cell lymphoma genomic DNA were subjected to PCR amplification using specific t(14;18) primers, which detected 100 pg of B-cell lymphoma DNA. PCR products (10 μl) were resolved on 8% PAGE/TBE gels and visualized after SYBR-Gold staining under UV illumination. F: Nested amplification of a PCR product from the Bcl-2 translocation t(14;18) gene from purified genomic DNA detected 10 pg of B-cell lymphoma DNA. PCR products (10 μl) were resolved on 8% PAGE/TBE gels and visualized after ethidium bromide staining under UV illumination. The Journal of Molecular Diagnostics 2007 9, DOI: ( /jmoldx ) Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

3 Figure 2 Seminested amplification of Fr2 PCR product generated from purified genomic DNA derived from patient samples. Positive control derived from DOHH genomic DNA. All PCR products (10 μl) were resolved on 8% PAGE/TBE gels and visualized after ethidium bromide staining under UV. The Journal of Molecular Diagnostics 2007 9, DOI: ( /jmoldx ) Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

4 Figure 3 Seminested amplification of Fr3 PCR product generated from purified genomic DNA derived from patient samples. Positive control derived from RAJI genomic DNA. All PCR products (10 μl) were resolved on 8% PAGE/TBE gels and visualized after ethidium bromide staining under UV. The Journal of Molecular Diagnostics 2007 9, DOI: ( /jmoldx ) Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

5 Figure 4 Nested amplification of PCR product from the Bcl-2 translocation t(14;18) gene from purified genomic DNA derived from patient samples. Positive control derived from DOHH genomic DNA. All PCR products (10 μl) were resolved on 8% PAGE/TBE gels and visualized after ethidium bromide staining under UV. The Journal of Molecular Diagnostics 2007 9, DOI: ( /jmoldx ) Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions


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