For analysis, 1 pg E. coli DH10B DNA was amplified with either the REPLI-g Single Cell DNA Library Kit or by the MALBAC (Multiple Annealing and Looping Based Amplification Cycles) method (Supplier Y) and sequenced on a MiSeq instrument (V2, 2x150 nt). The REPLI-g Single Cell DNA Library Kit generated >10x the yield, with significantly improved accuracy than the MALBAC method and ensured more even coverage than MALBAC.
For analysis, 1 cell, 10 cells or gDNA were used as starting material. WGA was carried out for 3 h or 8 h. The complexity of libraries prepared using the REPLI-g Single Cell DNA Library Kit was very high, as indicated by the extremely low percentage of duplicates detected, enabling efficient use of sequencing capacity. Similar results were obtained independent of the amount of starting material and incubation time.
Compared to a product from Supplier R, libraries generated using the REPLI-g Single Cell DNA Library Kit provided more uniform coverage over a wide range of GC content (%).
Data from libraries constructed from one HeLaS3 cell is shown. Compared to a PCR-based single cell library kit, the REPLI-g Single Cell DNA Library Kit provided a higher percentage of mapped reads and an extremely low percentage of duplicates.
The REPLI-g Single Cell DNA Library Kit provides a complete workflow for highly uniform amplification across the entire genome, with minimal sequence bias, followed by fast, one-tube library construction.