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Example sentences for: aca
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The HCMV DNA was detected using HCMV1 (5' cct agt gtg gat gac cta cgg gcc a) and HCMV2 (5' cag aca cag tgt cct ccc gct cct c) primers producing 249 bp long amplicon and the DNA of HHV6 was amplified with specific primer pair HP0 (5' ccg caa tcg aat cca cct agc gg) and HP4 (5' gtg aga acg gat tcg aac agt gct g) yielding 440 bp product [ 23 24 ] . All amplifications were carried out with 20 pmol of each primer in 2 mM solution of MgCl 2 , 2 U of Tag Special DNA polymerase (Biovendor, Czech Republic), 0.3 mM of each dNTPs, 10× reaction buffer and 1 μg of isolated DNA according to the following conditions: 96°C for 4 min, (94°C for 10 sec, 58°C for 10 sec, 72°C for 20 sec) 36 times, and final extension at 72°C for 2 min.
Replacement of the endogenous rec12 +locus with the rec12-Y98F allele was achieved by transformation and a pop-in, pop-out approach [ 61 63 ] . A 1.2 kbp Xba I- Xba I fragment of rec12 +was subcloned into pBluescript SKII(+) and subject to oligonucleotide directed mutagenesis [ 64 ] to simultaneously introduce a silent mutation generating an Eco RV site and to mutate the Y98 codon (GAG ATA TTT ATT ACA → GA G ATA T
A full length, 1.2 kb IDO cDNA was amplified from IFN-γ stimulated RAW cells and cloned into pGEM T-Easy (Promega), using primers; 5' TAG CGG CCG CGT AGA CAG CAA TGG CAC TC 3' forward, 5' TAA GAT CTT ACA CTA AGG CCA ACT CAG 3' reverse, which contain Not I and Bgl II sites respectively.
CT primer: GCA GGA GGG ACA GTT GAA GGA G
The primers for rat constitutively expressed ribosomal S12, were sense:5'-ACG TCA ACA CTG CTC TAC A-3' and antisense:5'-CTT TGC CAT AGT CCT TAA C-3' and generate a 303 bp cDNA product as previously described [ 63 ] . Controls for RT-PCR included samples without the enzyme reverse transcriptase and samples without template.
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