Optimized whole-genome amplification strategy for extremely AT-biased template.
Bottom Line: We present a strategy for whole-genome amplification (WGA) of low-yield samples from P. falciparum prior to short-read sequencing.We have developed WGA conditions that incorporate tetramethylammonium chloride for improved amplification and coverage of AT-rich regions of the genome.Our data show that this method is suitable for as low as 10 pg input DNA, and offers the possibility of sequencing the parasite genome from small blood samples.
Affiliation: Wellcome Trust Sanger Institute, Hinxton, UK email@example.com.Show MeSH
Related in: MedlinePlus
Mentions: Formation of chimeric reads is a major problem with MDA technology.27 Chimeras cause serious mapping and assembly problems, and therefore reduce the quality and quantity of the WGA product. We analysed the formation of chimeras by comparing WGA products following standard and optimized procedures. The number of chimeric reads increased as the amount of input DNA was reduced (Fig. 4). However, optimizing WGA condition by including TMAC additive decreased the formation of chimeras and improved the quality of reads in P. falciparum WGA.Figure 4.
Affiliation: Wellcome Trust Sanger Institute, Hinxton, UK firstname.lastname@example.org.