Single Cell Whole Genome Amplication Kit

MALBAC Single Cell Amp

• Why studying single cell?
• Multiple Annealing and Looping-based Amplification Cycles (MALBAC)
• Applications of Single Cell WGA kit
• Additional Information About MALBAC
• Ordering

Why studying single cell?

Cell-to-cell variability is universal in any organism. Even in cells with an identical genome, fluctuations of gene regulation and stochastic gene expression can cause significant deviation of individual cells from the population average. The information we get from a population average does not represent the single cell. Single cell analysis provides great insights into tumor heterogenity, stochastic gene expression and signaling.

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Multiple Annealing and Looping-based Amplification Cycles (MALBAC)

Nucleic acid analytical platforms (e.g. qPCR, microarray and high throughput sequencing) often require the sample to meet specific requirements for quantity and quality. Therefore, Whole genome amplification (WGA) is often required for analyzing samples with limited quantity, such as single circulating tumor cells, micro-biopsies, and single blastomere.

WGA methods are in general prone to amplification bias, PCR-based WGA introduces sequence-dependent bias mainly due to the exponential amplification. Multiple Displacement Amplification (MDA), which uses random priming and the strand-displacing phi29 polymerase under isothermal condition, has provided improvements over PCR-based methods but still exhibits considerable bias, again due to nonlinear amplification.

The Single Cell WGA kit generates highly uniform amplification across the entire genome. The method is based on the MALBAC (Multiple Annealing and Looping Based Amplification Cycles) technology *(1), which carries out close-to-linear pre-amplification cycles of the entire genome using a mixture of highly-processive DNA polymerases with strand displacement activity, followed by an exponential amplification by PCR to a sufficient amount for various downstream analyses.

malbac procedure

Figure 1: MALBAC procedure

The Single Cell WGA kit exhibits significantly improved amplification evenness compared with MDA. With 30x sequencing depth, 93% of the single cell genome can be covered compared with ~50-60% by MDA.

malbac vs mda

Figure 2: Comparison of uniformity of Single Cell Amplification between MALBAC and MDA

malbac vs mda

Figure 3: MALBAC generates significantly improved amplification evenness compared with MDA

Reference: (1) C. Zong, S. Lu, A.R. Chapman, X.S. Xie. Genome-Wide Detection of Single Nucleotide and Copy Number Variations of a Single Human Cell. Science. 338, 1622-1626 (2012)

malbac single cell WGA kit

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Applications of Single Cell WGA kit (from Yikon Genomics)

DNA amplified with the Single Cell WGA Kit can be used in a variety of downstream analytical platforms:

  • Detection of mutations  
  • SNP genotyping  
  • CNV profiling and aneuploidy detection
  • Whole genome and exome sequencing
  • Real-time PCR
  • aCGH microarray
  • Molecular cloning

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Additional Information About MALBAC Single Cell Analysis

Read more about MALBAC technologies for Single Cell Analysis from the articles below

• Zhao, Roseanne F."The Power of Sequencing Single Cell Genomes" National Human Genome Research Institute(January 22, 2013).
• Zakaib, Gwyneth Dickey. "New Method to Boost Single Cell Genomics" Alzheimer Research Forum (January 2013).
• Reuell, Peter. "One cell is all you need" Harvard Gazette (January 2013).
• Arnaud, Celia Henry. "Whole-Genome Sequencing Of Single Cells" Chemical & Engineering News (December 2012).
• Yeager, Ashley. "Single-Serve Sequencing: Sperm, Cancer" BioTechniques (December 2012).
• Karow, Julia. "Harvard Group Develops More Uniform Amplification Method for Single-Cell Sequencing" Genome Web(December 2012).

 

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Ordering

Product Size Cat #
Single Cell WGA Kit (Yikon Genomics) 10 reactions YK001A
Single Cell WGA Kit (Yikon Genomics) 50 reactions YK001B
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