|
|
Blood, Vol. 110, Issue 2, 578-586, July 15, 2007
A subpopulation of human peripheral blood NK cells that lacks inhibitory receptors for self-MHC is developmentally immature
Blood Cooley et al.
110: 578
Supplemental materials for Cooley et al, Vol 110, Issue 2, 578-586
Files in this Data Supplement:
- Figure S1. Dissociation curves of the amplified KIR products (PDF, 320 KB) -
Dissociation curves with SYBR green were generated by adding a melt program to the regular program consisting of 15 seconds at 95°C, 1 min at 60°C and 15 seconds at 95°C. SYBR Green binds double-stranded DNA non-specifically, labeling both amplicon and primer-dimers. Shown are dissociation/melting curves of the amplified products plotted as the negative derivative of fluorescence versus temperature (–dF/dT) against temperature for 2DS2, 2DL4, 2DL3 and 2DL2. KIR amplicons had melting temperatures of 80°C to 84°C (solid arrow), while primer-dimers had characteristically lower melting temperatures of 75°C (dashed arrow). KIR2DS2 and 2DL4 reactions amplified product but 2DL3 and 2DL2 reactions resulted in false readings due to primer-dimer formation.
- Figure S2. Standard curves for amplification of KIR genes (PDF, 89 KB) -
Standard curves of the amplification of all KIRs were performed using TaqMan probes and triplicate reactions with limiting dilutions of RNA (1000 to 0.0128 ng). Shown are representative standard curves for 2DS2, 2DL4, 2DL3 and 2DL2. The PCR efficiency was calculated using the equation E = 10–1/slope such that an ideal slope of -3.3 represents 100% efficiency of PCR reaction.
|
|
