Biotechnology Bulletin ›› 2021, Vol. 37 ›› Issue (10): 257-265.doi: 10.13560/j.cnki.biotech.bull.1985.2020-1478
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LIU Ying(), CHENG Li, ZHANG Bo()
Received:
2020-12-01
Online:
2021-10-26
Published:
2021-11-12
Contact:
ZHANG Bo
E-mail:liuy63@sustech.edu.cn;zhangb3@sustech.edu.cn
LIU Ying, CHENG Li, ZHANG Bo. High-resolution DNA Screening Based on Novel Magnetic Separation Technology[J]. Biotechnology Bulletin, 2021, 37(10): 257-265.
引物名称Primer | 引物序列Sequence |
---|---|
Y染色体正向引物(qPCR) | CATTCTCAAGCAAAACATGG |
Y染色体反向引物(qPCR) | CAGCAGTAGAGCAGTCAG |
Y染色体探针(qPCR) | CGTTGACTACTTGCCCT |
Y染色体正向引物(ddPCR) | CATTCTCAAGCAAAACATGG |
Y染色体反向引物(ddPCR) | CAGCAGTAGAGCAGTCAG |
Y染色体探针(ddPCR) | CGTTGACTACTTGCCCT |
X染色体正向引物(ddPCR) | AGTCACACCCACTTGTT |
X染色体反向引物(ddPCR) | GGGCCATACAATCTGTTG |
X染色体探针(ddPCR) | CTCTGACCTGGTAGCAC |
Table 1 Information of primers and probes
引物名称Primer | 引物序列Sequence |
---|---|
Y染色体正向引物(qPCR) | CATTCTCAAGCAAAACATGG |
Y染色体反向引物(qPCR) | CAGCAGTAGAGCAGTCAG |
Y染色体探针(qPCR) | CGTTGACTACTTGCCCT |
Y染色体正向引物(ddPCR) | CATTCTCAAGCAAAACATGG |
Y染色体反向引物(ddPCR) | CAGCAGTAGAGCAGTCAG |
Y染色体探针(ddPCR) | CGTTGACTACTTGCCCT |
X染色体正向引物(ddPCR) | AGTCACACCCACTTGTT |
X染色体反向引物(ddPCR) | GGGCCATACAATCTGTTG |
X染色体探针(ddPCR) | CTCTGACCTGGTAGCAC |
Fig.2 Verification of short DNA fragments enrichment A: DNA fragments distribution before and after enrichment. B: DNA fragments distribution(below 150 bp)before and after enrichment. C: Average length of cfDNA main peak before and after entichment
Fig.3 Enrichment efficiency evaluation by ddPCR A: Workflow of copy number calculation for ChrX and ChrY by ddPCR before and after enrichment. B: Fluorescence signal of sample before and after enrichment. C: ChrY% statistics of 21 samples from pregnant women with male fetus. D: ChrY% fold change for 21 samples from pregnant women with male fetus
Fig. 5 Enrichment of short cfDNA fragments with fetal bovine serum by optimized workflow A: Changes in DNA fragments distribution before and after 2-step selection enrichment. B: Fragment distribution by 2-time selection and 2-step long fragments DNA separation with the peak beginning position at 150 bp. C: Proportion of DNA below 150 bp before and after enrichment. D: Average length change of cfDNA main peak
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