I used my own SNP (pronounced 'snip') as a seed for the algorithm on which each work is structured; I identified twenty-three variables that were then put through a random function to generate the set of instructions that I followed when knitting".
The authors used an iterative process of drafting, evaluation, and revision to generate the items, response options, instructions, and recall period for the instrument.
A post-processor then runs through these instructions to generate the segment identities.
The PAS Entry clone for ABACUS1 was altered using QuikChange (Agilent, Santa Clara, CA) site-directed mutagenesis according to manufacturer's instructions to generate the ABACUS affinity mutants.
AccuTaq LA DNA polymerase (Sigma) was used in accordance with the manufacturer's instructions to generate the DNA fragments for cloning and integration.
Standard oligos containing the 5′ T7 RNA polymerase recognition sequence were designed for these vectors and used with the Megascript kit (Ambion) according to manufacturer's instructions to generate the dsRNAs used for the transfections.
The Stratagene QuikChange Mutagenesis kit was used according to manufacturer's instructions to generate the moderate mutator dnaQ928, which causes the G17S substitution (Taft-Benz and Schaaper 1998) in the context of plasmid pMM5Z.
Quick-change II XL Site-Directed Mutagenesis kit (Stratagene) was used according to the manufacturer's instructions to generate the mutations of putative PKC phosphorylation sites within the regulatory C-terminal domain of HCN2: T549A, T566A, T566A, S818A.
Five nanograms of total RNA with an RNA integrity number > 8 were amplified using Ovation Pico WTA System V2 (NuGEN, San Carlo, CA, USA) according to the manufacturer's instructions, to generate the Single Primer Isothermal Amplification (SPIA) cDNA.
The mylz2 GFP plasmid was also used as a template for in vitro mutagenesis of the five sites containing the Prdm1 GAAAG core sequence using the QuikChange® Multi Site-Directed Mutagenesis Kit (Stratagene, La Jolla, CA, USA) following the manufacturer's instructions to generate the Tg((5M mylz2 GFP i130 transgenic line.
