TACI Expression and Signaling in Chronic Lymphocytic Leukemia

Table 1

Primers and PCR conditions for the amplification of the analyzed genes.

Gene	Primers	Sequence	PCR conditions
TNFRSF13B	Forward; Reverse	5′-TCTGCCTGTGTGCCGTCCTC-3′; 5′-CGGCTTCCATCGCGTGAT-3′	95°C for 2 min, followed by 40 cycles (95°C for 10 s, 64°C for 60 s)
B2M	Forward; Reverse	Commercially obtained by Qiagen, Cat. No PPH01094E	95°C for 10 min, followed by 40 cycles (95°C for 15 s, 60°C for 60 s)
IGHV; A′ PCR	FR2A; FR1C; LGH	5′-TGG(A/G)TCCG(C/A)CAG(G/C)C(T/C)(T/C)C(A/G/T/C)-3′; 5′-AGGTGCAGCTG(C/G)(A/T)G(CG)AGTC(A/G/T)GG-3′; 5′-TGAGGAGACGGTGACC-3′	95°C for 2 min, followed by 5 cycles (95°C for 30 s, 63°C for 30 s, 72°C for 30 s), then by 25 cycles (95°C for 30 s, 57°C for 30 s, 72°C for 30 s), and a final elongation at 72°C for 5 min
IGHV; nested PCR	FR2A; VH1; VH2; VH3; VH4; VLJH	Similar with A′ PCR; 5′-ACTAGTCGACCTCAGTGAAGGT-3′; 5′-ACTAGTCGACGTCCTGVGCTGGTGAAA(G/C)CCACAC-3′; 5′-ACTAGTGACGGGTCCCTGAGACTCTCCTGTGCAG-3′; 5′-ACTAGTCGACCCTGTCCCTCACCTGC(A/G)CTGTC-3′; 5′-GTGACCAGGGT(A/G/C/T)CCTTGGCCCCAG-3′	95°C for 2 min, followed by 30 cycles (95°C for 30 s, 63°C for 30 s, 72°C for 30 s), and a final elongation at 72°C for 5 min

B2M: beta-2-microglobulin; IGHV: immunoglobulin heavy chain variable region.