MALBAC

Prior to MALBAC, a single cell is isolated by various methods including laser capture microdissection, microfluidic devices, flow cytometry, or micro pipetting, then lysed.

MALBAC single-cell whole-genome amplification involves 5 cycles of quenching, extending, melting, and looping.

The utilization of specialized primers enables looping of amplicons which then prevents them from being further amplified in subsequent cycles of MALBAC.

MALBAC may aid in the analysis of forensic specimens, in pre-natal screening for genetic diseases, in understanding the development of reproductive cells, or in elucidating the complexity of a tumour.

It may be used to examine intratumor heterogeneity, to identify genes which may confer an aggressive or metastatic phenotype, or to evaluate the potential for a tumour to develop drug resistance.

[4][5] A pioneering application of MALBAC was published in a December 2012 issue of Science and described the use of this technology to measure the mutation rate of the colon cancer cell line SW4802.

By sequencing the genomes of 99 individual human sperm cells from an anonymous donor, MALBAC was used to examine genetic recombination events involving single gametes and ultimately provide insight into the dynamics of genetic recombination and its contribution to male infertility.

[6] Additionally, within an individual sperm, MALBAC identified duplicated or missing chromosomes, as well as SNPs or CNVs which could negatively affect fertility.

[4][6] MALBAC is a form of whole genome sequencing which reduces the bias associated with exponential PCR amplification by using a quasilinear phase of pre-amplification.

[4][7] Only a small amount of starting template (picograms of DNA) is required to initiate the process, and therefore it is an ideal method for the sequencing of a single human cell.