Single-cell genomic analysis has grown rapidly in recent years and will find widespread applications in various fields of biology, including cancer biology, development, immunology, pre-implantation genetic diagnosis, and neurobiology. In this study, we amplified genomic DNA from individual hippocampal neurons using one of three single-cell DNA amplification methods (multiple annealing and looping-based amplification cycles (MALBAC), multiple displacement amplification (MDA), and GenomePlex whole genome amplification (WGA4)). We then systematically evaluated the genome coverage, GC-bias, reproducibility, and copy number variations among individual neurons. Our results showed that single-cell genome sequencing results obtained from the MALBAC and WGA4 methods are highly reproducible and have a high success rate. Chromosome-level and subchromosomal-level copy number variations among individual neurons can be detected.
Since the publication of this paper the authors have noticed there were errors in two series of letters in the Materials and Methods section. The correct paragraph is shown below. The authors would like to apologize for any inconvenience caused.Expression and purification of TAT-N24 and control TAT-N24M fusion proteins. cDNA fragments encoding 6xHis tag, PDT in Tat protein (YGRKKRRQRRR), HA tag (YPYDVPDYA) and N24 (MDRDDADWREVMMPYSTELI-FYIE) were introduced into pMD18-T Easy vector (Invitrogen, Grand Island, NY, USA). His-TAT-HA-N24 cDNA fragment then was subcloned into the vector pET32a (Invitrogen). To generate the control fusion protein (TAT-N24M), the N24 cDNA sequence was replaced by cDNA encoding the shuffled N24 (MDRDDWDAREVIMMPYSELYTIFE). The TAT-N24 and TAT-N24M fusion proteins have 96 amino-acid residues. The obtained recombinant vectors (designated pHisTAT-N24 and pHisTAT-N24M) were transformed into E. coli BL21 (DE3) for fusion protein expression.
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