Comparison of different lysis buffers in the assessment of the efficiency of single cell polymerase chain reaction for preimplantation genetic diagnosis
Li, Q.; Ma, Y.-l.; Pan, Q.
Hunan Yi Ke da Xue Xue Bao 27(3): 201-203
2002
ISSN/ISBN: 1000-5625 PMID: 12575290 Document Number: 551946
To set up a highly efficient cell lysis system to increase the efficiency of single cell polymerase chain reaction (PCR) for preimplantation genetic diagnosis (PGD). First we picked up one single lymphocyte or blastomere and lysed it in alkaline lysis buffer (ALB), proteinase K buffer (PK) and a new proteinase K lysis buffer compounded by ourselves. Then we amplified the 3rd exon of Gene ALD using nested PCR and compared the amplification efficiencies. The amplification efficiencies of the single lymphocyte in ALB, PK and the lysis buffer compounded by ourselves were 90%, 74%, and 87%; those of the single blastomere were 98%, 77%, and 96%. ALB and the lysis buffer compounded by ourselves are superior to PK in the lysis of cells in clinical PGD.