Manjusha Dixit, Department of Medical Genetics, Sajay Gandhi Post Graduate Institute of Medical Sciences, Lucknow, India
3 February 2004
We read paper by Chhabra et al. (1) and found it very interesting but we would like to draw the attention of authors towards some points. Firstly, primer sequences used for amplification of desired region of Apolipoprotein CIII gene does not match with the sequences given in reference article (2). Sequences reported in the paper are, forward primer: 5'- CAT GGT TGC CTA CAG GAG TTC-3' and reverse: 3'-TGA CCT TCC GCA CAA AGC TGT-5', but in reference article reverse primer is 5'-TGA CCT TCC GCA CAA AGC TGT-3'. Secondly, sequence of forward primer does not show 100% match with any of the Apo CIII gene entries in NCBI database. For example with a sequence " accession XO1392.1 GI:28725", only bases 1-15 of forward primer match with base 2955-2969 and bases 16-21 match with base 2972-2977. In other words, two bases between 15-16 of the forward primer are missing. Thirdly, sequence alignment of forward and reverse primer using BLAST shows that amplified product will be 591 bp long instead of 596 bp as reported by Chhabra et al.(1).
1. Chhabra S, Narang R, Krishnan LR, Vasisht, Agarwal DP, Srivastava LM, Manchanda SC, Das N. Apolipoprotein C3 SstI polymorphism and triglyceride levels in Asian Indians. BMC Genetics 2002:3: 9.
2. Paul-Hayase H, Rossenue M, Robinson D, van Bervliet JP, Deslypere JP, Humphries SE: Polymorphism in the apolipoprotein (apo) AI-CIII-AIV gene cluster: detection of genetic variation determining plasma apo AI, apo CIII, and apo AI concentrations. Hum Genet 1992, 88:439-446.
Discrepancy in PCR methodology
3 February 2004
We read paper by Chhabra et al. (1) and found it very interesting but we would like to draw the attention of authors towards some points. Firstly, primer sequences used for amplification of desired region of Apolipoprotein CIII gene does not match with the sequences given in reference article (2). Sequences reported in the paper are, forward primer: 5'- CAT GGT TGC CTA CAG GAG TTC-3' and reverse: 3'-TGA CCT TCC GCA CAA AGC TGT-5', but in reference article reverse primer is 5'-TGA CCT TCC GCA CAA AGC TGT-3'. Secondly, sequence of forward primer does not show 100% match with any of the Apo CIII gene entries in NCBI database. For example with a sequence " accession XO1392.1 GI:28725", only bases 1-15 of forward primer match with base 2955-2969 and bases 16-21 match with base 2972-2977. In other words, two bases between 15-16 of the forward primer are missing. Thirdly, sequence alignment of forward and reverse primer using BLAST shows that amplified product will be 591 bp long instead of 596 bp as reported by Chhabra et al.(1).
1. Chhabra S, Narang R, Krishnan LR, Vasisht, Agarwal DP, Srivastava LM, Manchanda SC, Das N. Apolipoprotein C3 SstI polymorphism and triglyceride levels in Asian Indians. BMC Genetics 2002:3: 9.
2. Paul-Hayase H, Rossenue M, Robinson D, van Bervliet JP, Deslypere JP, Humphries SE: Polymorphism in the apolipoprotein (apo) AI-CIII-AIV gene cluster: detection of genetic variation determining plasma apo AI, apo CIII, and apo AI concentrations. Hum Genet 1992, 88:439-446.
Competing interests
None declared