Some of them corresponded to silent mutations (GTC to GTT at V74; ATC to ATT at I65; GAC to GAT at D39, and TGA to TAA at the stop codon of the gene).
Fig. 9 Plot of the linear absorption coefficient at different GAC layers in the rum filter and the GAC-virgin.
The plot of the linear absorption coefficient at different GAC layers in the rum filter and the GAC-virgin is presented in Fig. 9.
Table 4 depicts the spatial concentration values at different GAC layers in the rum filter taking the GAC-Top as GSI reference.
We also amplified DNA fragments for 9 DNA samples (3 each of C/C, C/T and C/T genotypes) previously genotyped by allele discrimination (forward primer: 5' TGC CTT TTA AAA TCC AAA GAC AT; reverse primer: 5' AGA GCG AGA CCA GGA ACA AA) spanning the rs3135005 C/T SNP [15].
Although evident differences between GAC-Top and GAC-virgin can be observed in terms of ( overline{mathrm{GSI}} ) and the linear absorption coefficient, no significant statistical differences in the ( overline{ mathrm{GSI}} ) and μ values for comparing the exhaustion level at different GAC layers in the rum filter were found.
The degenerate primers used were E23P1 (forward primer; 5′-CCG YAG GTA ACA AGW GAC AT-3′) (5 ) and 35R (reverse primer; 5′-TCT CAG CAC TAA TGA CCC TC-3′).
For the screening experiments, further Bcl-xL antisense oligonucleotides were tested: ISIS 15999 (5′-TCC CGG TTG CTC TGA GAC AT-3′), ISIS 16011 (5′-CTG GAT CCA AGG CTC TAG GT-3′), and ISIS 22783 (5′-CTG GAT CCA AGG CTC TAG GT-3′).
The reaction included TaqMan universal master mix (Applied Biosystems), CMV –gB-F: 5′-TGG GCG AGG ACA ACG AAA TC-3′, CMV –gB-R: 5′-TGA GGC TGG GAA GCT GAC AT-3′ as primer and 5′ FAM) -TGG GCA ACC ACC GCA CTG AGG – (BHQ) 3′ as probe (modified, [ 35]), respectively.
Briefly, we amplified a short PCR fragment containing a diagnostic SNP (eyeF, 5′-TGT GTG AGC AAA ATT CTC GG-3′; eyeR, 5′-GTT TCG GCA TGG TAG GAC AT-3′), digested with MboII, and genotyped by separating restriction fragments on a 2.5% agarose gel.
