The eno genes were amplified from chromosomal DNA using forward and reverse primers incorporating a BamHI or SalI restriction site, respectively (primer 1: GCG GAT CCA TGT CAA TTA TTA CTG ATG TTT ACG, primer 2: GCT GTC GAC TTA TTA TTT TTT AAG GTT GTA GAA TGA TTT).
This 'learning to reverse' might incorporate other neuroanatomical structures that are not specific to reversal learning, hence the lack of commonality in reversal loading.
Forward and reverse primers incorporated a HindIII and a BamHI, respectively.
To create the lsk1-K306R HA tagged mutant, A N-terminal fragment of the lsk1 gene was PCR amplified using primer JK6 (5'-ggg ggg aat tcg aac agc gga aaa gaa cat tt-3') and a reverse primer incorporating the K306R mutation, JK47 (5'-cct tat ccg tcg taa agc tac-3').
Two further fluorophores are used to make a sequencing ladder, either by reverse transcription, incorporating one ddNTP, or by cycle-sequencing.
A 1.5-kb 5′-homology arm leading up to the S1 cleavage site was amplified (Phusion polymerase, New England Biolabs) with a forward primer containing a SpeI site and a reverse primer incorporating a partial (up to the XbaI site) FRT site.
The mouse CAR ligand-binding domain from amino acids 118 358 (mCAR-LBD) was amplified using 5′-GCGGAATTCGGTACTAAAGCATCTTTGCAACTGAA-3′ and 5′-GCGGATCCTCAACTGCAAATCTCCCCGAGC-3′ as forward and reverse primers respectively, incorporating 5′ EcoRI and 3′ BamHI sites to facilitate cloning.
For each sample to be analyzed by microarray, 4 μg amplified RNA were primed with 5 μg random hexamers and 5 μg oligo dT and reverse-transcribed, incorporating aa-dUTP, as described at.
When the field strengths is greater than 3 T, large peak fields produced between adjacent coils with alternating currents limit the ability to incorporate reverse current coils without a breach of the peak field limit.
When screening a limited number of genes, it is more advantageous to use gene specific primers with the forward and reverse adapters already incorporated in the oligo's.
