ABCMdb

PMID: 15858148

Jarvis M, Iyer RK, Williams LO, Noll WW, Thomas K, Telatar M, Grody WW
A novel method for creating artificial mutant samples for performance evaluation and quality control in clinical molecular genetics.
J Mol Diagn. 2005 May;7(2):247-51., [PubMed]

Sentences

No. Mutations Sentence Comment

5 ABCC7 p.Gly85Glu CFTR mutations G85E and 1078delT were chosen to represent mutations in the original American College of Medical Genetics-recommended population- screening panel of 25 mutations. Login to comment 37 ABCC7 p.Asn1303Lys ABCC7 p.Gly85Glu It is ϳ250 kb in size and contains 27 exons.8 Several mutations were chosen for preliminary targeting experiments, including G85E (exon 3), N1303K (exon 21), and 1078delT (exon 7). Login to comment 39 ABCC7 p.Asn1303Lys ABCC7 p.Gly85Glu The study described here focused on the G85E and 1078delT target sequences because they are rarer and harder to obtain from natural sources than N1303K; the latter was used as a marker primarily to ensure that our CFTR constructs potentially encompassed all possible mutations in the original ACMG panel. Login to comment 60 ABCC7 p.Gly85Glu ABCC7 p.Gly85Glu Because it was the major DNA component, we determined that when salmon sperm DNA was used as template for PCR analysis using any of our usual oligonucleotide primer sets, no amplified products of the anticipated sizes were observed by either an in-house amplification restriction digestion method or a commercial CFTR hybridization assay (Roche Diagnostics, Indianapolis, IN).10 Samples were formulated and analyzed for each of the following five genotypes: wild-type (homozygous normal), homozygous G85E, homozygous 1078delT, heterozygous G85E, and heterozygous 1078delT. Login to comment 62 ABCC7 p.Gly85Glu Results of in-house pilot testing of constructed heterozygous and homozygous products for CFTR mutations G85E and 1078delT using a commercial reverse hybridization strip system (Roche Linear Array CF Gold 1.0). Login to comment 64 ABCC7 p.Gly85Glu ABCC7 p.Gly85Glu The observed genotypes are: lane 1, negative for tested mutations; lane 2, G85E homozygote; lane 3, 1078delT homozygote; lane 4, G85E heterozygote; lane 5, 1078delT heterozygote; lane 6, negative for tested mutations. Login to comment 93 ABCC7 p.Gly85Glu ABCC7 p.Gly85Glu Pilot Testing Results Sample Laboratory analysis (correct results/total results) Unable to analyze Normal (w.t.) 7/8 1 G85E, heterozygous 8/9 G85E, homozygous 8/9 1078delT, heterozygous 8/9 1078delT, homozygous 7/9 Table 2. Login to comment