OBJECTIVE: Pre-implantational genetic diagnosis (PGD) from polar bodies or blastomeres has to be performed on single cells after whole genome amplification without allele drop out (ADO). Recently, an isothermal DNA amplification method, termed multiple displacement amplification (MDA) has been applied to single cells, leading to the synthesis of DNA with limited sequence representation bias. Thus, we aimed to determine the efficacy of MDA in amplifying the whole genomic DNA from polar bodies and single blastomeres intended for subsequent sequence tandem repeat amplification by polymerase chain reaction (STR-PCR) as performed for single gene disorders testing.DESIGN: Prospective laboratory study.MATERIALS AND METHODS: After biopsy, polar bodies (n=8) and blastomeres (n=8) were individually transferred to tubes containing 2μL of ultra-pure water and submitted to drastic temperature changes by immersion in N2L for cell rupture. This material was subjected to the MDA protocol by addition of non-specific primers and DNA polymerase phi29 following the manufacturers' instruction (REPLI-g Mini Kit, Qiagen Inc., USA). After the isothermal amplification, DNA was quantified by spectrophotometry and an aliquot of 0.5 ng/μL was transferred to another tube containing the PCR master mix for amplification of 15 sequence tandem repeats (STR) (loci: Penta E, D18S51, D21S11, TH01, D3S1358, FGA, TPOX, D8S1179, vWA, Amelogenin, Penta D, CSF1PO, D16S539, D7S820, D13S317 and D5S818) and amelogenin for 32 thermal cycles. STR peaks were detected in an automatic DNA sequencer. Allele drop out was estimated by another two subsequent MDAs, each one followed by a new STR analysis for comparison to result obtained from the first MDA/STR-PCR.RESULTS: DNA concentrations after the first MDA of whole genome of second polar bodies and blastomeres were 1.7±0.27 μg of DNA/ μL (mean ± standard error) and 3.8±0.42 μg of DNA/ μL. Analysis of PCR amplifications of STR resulted in 14.25±0.12 peaks after the first MDA. The second MDA/STR-PCR produced 12.37±0.18 peaks and the third MDA/STR-PCR produced 10.5±0.15 peaks. The mean allele drop out was 14.15%.CONCLUSIONS: Multiple displacement amplification is an efficient technique for amplification of the whole genome of second polar bodies and single blastomeres and the resulting amplicons can be subjected to multiple downstream applications such as, DNA sequencing, short tandem repeat analysis, and mutation analysis. OBJECTIVE: Pre-implantational genetic diagnosis (PGD) from polar bodies or blastomeres has to be performed on single cells after whole genome amplification without allele drop out (ADO). Recently, an isothermal DNA amplification method, termed multiple displacement amplification (MDA) has been applied to single cells, leading to the synthesis of DNA with limited sequence representation bias. Thus, we aimed to determine the efficacy of MDA in amplifying the whole genomic DNA from polar bodies and single blastomeres intended for subsequent sequence tandem repeat amplification by polymerase chain reaction (STR-PCR) as performed for single gene disorders testing. DESIGN: Prospective laboratory study. MATERIALS AND METHODS: After biopsy, polar bodies (n=8) and blastomeres (n=8) were individually transferred to tubes containing 2μL of ultra-pure water and submitted to drastic temperature changes by immersion in N2L for cell rupture. This material was subjected to the MDA protocol by addition of non-specific primers and DNA polymerase phi29 following the manufacturers' instruction (REPLI-g Mini Kit, Qiagen Inc., USA). After the isothermal amplification, DNA was quantified by spectrophotometry and an aliquot of 0.5 ng/μL was transferred to another tube containing the PCR master mix for amplification of 15 sequence tandem repeats (STR) (loci: Penta E, D18S51, D21S11, TH01, D3S1358, FGA, TPOX, D8S1179, vWA, Amelogenin, Penta D, CSF1PO, D16S539, D7S820, D13S317 and D5S818) and amelogenin for 32 thermal cycles. STR peaks were detected in an automatic DNA sequencer. Allele drop out was estimated by another two subsequent MDAs, each one followed by a new STR analysis for comparison to result obtained from the first MDA/STR-PCR. RESULTS: DNA concentrations after the first MDA of whole genome of second polar bodies and blastomeres were 1.7±0.27 μg of DNA/ μL (mean ± standard error) and 3.8±0.42 μg of DNA/ μL. Analysis of PCR amplifications of STR resulted in 14.25±0.12 peaks after the first MDA. The second MDA/STR-PCR produced 12.37±0.18 peaks and the third MDA/STR-PCR produced 10.5±0.15 peaks. The mean allele drop out was 14.15%. CONCLUSIONS: Multiple displacement amplification is an efficient technique for amplification of the whole genome of second polar bodies and single blastomeres and the resulting amplicons can be subjected to multiple downstream applications such as, DNA sequencing, short tandem repeat analysis, and mutation analysis.