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. 2019 Jun;5(3):a004101. doi: 10.1101/mcs.a004101

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© 2019 Okur et al.; Published by Cold Spring Harbor Laboratory Press

This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted reuse and redistribution provided that the original author and source are credited.

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Figure 2. — Two percent agarose-gel electrophoresis image of the cDNA studies performed with two sets of primers. Peripheral blood samples were collected from one affected individual (II-6), unaffected carrier parents (I-2 and I-1), and a control. (A) Two different sets of primers were designed, given the lack of exon 3 in one noncoding isoform (NR_045492.1). Forward primer of set 1 (LSM1splice1F; yellow-filled, red outlined right arrow) maps to first exons of NM_014462.2 and NR_045492.1 only. Forward primer of set 2 (LSM1splice2F; yellow-filled, black outlined right arrow) maps to exon 3 of NM_014462.2 and NR_045493.1 only. Reverse primers of both sets (LSM1splice1R and LSM1splice2R) map to exon 4 of all isoforms. Isoform accession numbers and schematic were retrieved from NCBI Entrez Gene website (Gene ID = 27257). (B) PCRs with primer sets 1 (left) and 2 (right) revealed no band in an affected individual (II-6) corresponding to the canonical isoform. Unaffected carrier parents (I-2 and I-1) only have one band corresponding to the canonical isoform, 250-bp band on the left and 176-bp band on the right, in addition to the 134-bp noncoding RNA transcript (NR_045492.1) product (smaller band on the left) in all samples run with primer set 1.