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Fig. 4 | Stem Cell Research & Therapy

Fig. 4

From: Revealing cellular and molecular complexity of the central nervous system using single cell sequencing

Fig. 4

Whole-genome amplification methods for single cell sequencing. a Degenerate oligonucleotide-primed PCR (DOP-PCR). The 3′ end of the degenerate oligonucleotide primer (the random six nucleotides) are annealed to the genomic template, allowing the primer to initiate PCR, and PCR fragments are generated to contain the full length of the oligonucleotide primer at one end and the complementary sequence at the other end. Subsequently, the temperature is increased to amplify the DNA fragments. b Multiple displacement amplification (MDA). Double-stranded DNA are melted and random primers are bound to the DNA strand. Branched structures are produced under isothermal condition using the phi29 DNA polymerase. c Multiple annealing and looping-based amplification cycles (MALBAC). Double-stranded DNA is denatured into single strands at 94 °C, then single-stranded DNA templates are homogeneously bound with random primers at 0 °C. Semi-amplified products (n) are further amplified to produce full amplicons during a subsequent five temperature cycles (m), and the complete amplification products are 5′ and 3′ complementary to each other. Cyclization of the complete amplification product is performed with the temperature dropped to 58 °C to prevent further amplification and hybridization of the sequence. Semi-amplification products and genomic DNA continue to circulate to generate the complete amplification product

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