Restriction sites BamH1 and EcoRI in primers An--F/R and the mutated site are underlined; Start and stop codons are marked in bold; site-directed mutagenesis site is underline in I249L-F and I249L-R.
Bold: sites at which influenza was detected.
Bold: sites where influenza was detected.
Additional sequence was added to encode a His-tag, EcoRI (underlined) and XbaI (bold) sites and an additional tail to enhance cloning.
The human H1 RNA promoter was PCR-amplified from p Silencer™ 3.1-H1 hygro (Ambion) with the following primers: 5′-GTGAATTCATATTTGCATGTCGCTATGTG-3′ and 5′-AAAAAGCTT CCAAAAAATTGGTT CCATGGGAATGGAGATCTGTGGTCCATACAGA-3′, to introduce one BglII and two BstXI (bold) sites.
dpn also contained a pair of sites that conforms to the SPS configuration frequently found in Su(H) enhancers (CSB sequence: AAT GTGAGAAAAAAACT TTCTCACGATCACCTT, Su(H) sites in bold, Pbx site is ATCA).
Folks who have been to the Bold Moves site spend 12% more time looking at cars, configuring them and searching dealer inventory than other visitors to the sales site.
This was followed by high fidelity PCR amplification using the reverse (RT primer) and forward (5'-AAA CCATGGTACTAATACGACTCACTAAA-3'; bold, NcoI site).
The final products were again gel purified by size fractionation and submitted to reverse transcription reaction using the RT primer (5'-TTTT CTGCAGAAGGATGCGGTTAAA-3'; bold, PstI site).
Sites under selection were predicted using the Bayes Empirical Bayes (BEB) approach: sites inferred to be under positive selection with posterior probabilities >95% are in bold and sites with posterior probabilities of > 99% are indicated by an asterisk.
