CRISPR systems and their associated nucleases are endonucleases that target DNA cut sites through small guide RNAs (gRNAs). gRNAs contain a variable 20-nucleotide sequence for targeting specific DNA cut sites (crRNA), and a longer scaffold sequence is required for nuclease binding (tracrRNA). For convenience, single-guide RNAs (sgRNAs) link the crRNA and tracrRNA into a chimeric molecule. CRISPR screens use libraries of sgRNA sequences delivered in vectors as DNA for in vivo transcription to gRNAs. Often the most challenging part of a screen is not its characterization, but the cloning of the oligo pool into a vector under an inducible promoter. The problems are two-fold: first is the lack of high quality oligo pools and the second is bias in oligo sequence representation after amplification, cloning, and growth. We highlight here the ability of Twist Bioscience’s multiplexed oligo pool libraries to maintain uniformity throughout amplification and cloning. We measure cloning method efficiency by the number of colony-forming units (CFUs) and measure sequence distributions and uniformity after amplification and cloning using next-generation sequencing (NGS). We demonstrate the correction of method-specific cloning biases with longer inserts for increased sequence uniformity in the final cloned pool.