Method for single cell genome analysis and kit therefor
A method for analyzing a genome of a single cell is provided, and a kit is also provided. The method for analyzing the genome of the single cell may comprise separating and lysing the single cell to obtain a whole-genome DNA of the cell; subjecting the whole-genome DNA to a whole-genome amplification to obtain a whole-genome amplification product; performing a PCR amplification using the whole-genome amplification product as template and using housekeeping-gene-specific primers to detect the housekeeping gene of the whole-genome amplification product; and determining whether the whole genome amplification product meets a requirement for sequencing based on the detection result, wherein a uniform distribution of the amplification product in each chromosome is an indication of the amplification product meeting the requirement for sequencing.
1. A method for analyzing a genome of a human single cell comprising the steps of:
separating and lysing the single cell to obtain a whole-genome DNA of the cell;
subjecting the whole-genome DNA to a whole-genome amplification to obtain a whole-genome amplification product;
performing a PCR amplification using the whole-genome amplification product as a template and using housekeeping-gene-specific primers to detect a set of housekeeping genes of the whole-genome amplification product, wherein the set comprises PRDX6, RPL37a, ADD1, HLA-A, RAD9A, ARHGEF7, EIF2B2, PSMD7, BCAT2 and ATP5O; and
determining whether the whole genome amplification product meets a requirement for sequencing based on the detection result,
wherein a uniform distribution of the amplification product in each chromosome is an indication of the amplification product meeting the requirement for sequencing.
2. A method according to claim 1 , wherein the whole-genome amplification comprises multi-displacement amplification (MDA) or DOP-PCR whole-genome amplification.
3. A method according to claim 1 , wherein
for PRDX6, the housekeeping-gene-specific primers have nucleotide sequences shown as SEQ ID NO:1 and SEQ ID NO:2, respectively;
for RPL37a, the housekeeping-gene-specific primers have nucleotide sequences shown as SEQ ID NO:3 and SEQ ID NO:4, respectively;
for ADD 1, the housekeeping-gene-specific primers have nucleotide sequences shown as SEQ ID NO:5 and SEQ ID NO:6, respectively;
for HLA-A, the housekeeping-gene-specific primers have nucleotide sequences shown as SEQ ID NO:7 and SEQ ID NO:8, respectively;
for RAD9A, the housekeeping-gene-specific primers have nucleotide sequences shown as SEQ ID NO:9 and SEQ ID NO:10, respectively;
for ARHGEF7, the housekeeping-gene-specific primers have nucleotide sequences shown as SEQ ID NO:11 and SEQ ID NO:12, respectively;
for EIF2B2, the housekeeping-gene-specific primers have nucleotide sequences shown as SEQ ID NO:13 and SEQ ID NO:14, respectively;
for PSMD7, the housekeeping-gene-specific primers have nucleotide sequences shown as SEQ ID NO:15 and SEQ ID NO:16, respectively;
for BCAT2, the housekeeping-gene-specific primers have nucleotide sequences shown as SEQ ID NO:17 and SEQ ID NO:18, respectively;
for ATP5O, the housekeeping-gene-specific primers have nucleotide sequences shown as SEQ ID NO:19 and SEQ ID NO:20, respectively.
4. A method according to claim 1 , wherein the whole-genome amplification product is subjected to housekeeping gene quantitative detection.
5. A method according to claim 1 , further comprising constructing a DNA sequence library of the amplification product, wherein the amplification product meets the requirement for sequencing.
6. A method according to claim 5 , further comprising sequencing the DNA sequence library.