American Society of Hematology

Fig. 4.

$Fig. 4. Representative seminested PCR analysis of H-RS cell-specific DNA in apheresis material, bone marrow, spleen, and lymph node samples from a patient with NS-HD (patient no. 1; A and B) and a patient with MC-HD (patient no. 2; C and D). DNA samples (1 μg) were amplified in five replicates (except control PBMCs for patient no. 1) using NSfor and NSrev1/rev2 primers (patient no. 1) and MCfor and MCrev1/rev2 primers (patient no. 2). PCR products were fractionated by agarose gel electrophoresis (2%) and stained with ethidium bromide (A and C). After Southern blot, the DNA was hybridized to the internal FITC-labeled oligonucleotide NSintern (B) or MCintern (D) and visualized by ECL. M, 100-bp DNA ladder; P, PCR product amplified from single H-RS cells served as the positive controls (NSVH1, 249 bp; MCVH1, 142 bp). Negative controls: DNA from a healthy volunteer (PBMC*), water control after first and second (#) PCR.$

Representative seminested PCR analysis of H-RS cell-specific DNA in apheresis material, bone marrow, spleen, and lymph node samples from a patient with NS-HD (patient no. 1; A and B) and a patient with MC-HD (patient no. 2; C and D). DNA samples (1 μg) were amplified in five replicates (except control PBMCs for patient no. 1) using NSfor and NSrev1/rev2 primers (patient no. 1) and MCfor and MCrev1/rev2 primers (patient no. 2). PCR products were fractionated by agarose gel electrophoresis (2%) and stained with ethidium bromide (A and C). After Southern blot, the DNA was hybridized to the internal FITC-labeled oligonucleotide NSintern (B) or MCintern (D) and visualized by ECL. M, 100-bp DNA ladder; P, PCR product amplified from single H-RS cells served as the positive controls (NSVH1, 249 bp; MCVH1, 142 bp). Negative controls: DNA from a healthy volunteer (PBMC*), water control after first and second (#) PCR.

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