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. 2016 May 5;11(5):e0154855. doi: 10.1371/journal.pone.0154855

Fig 7. Cloning of a mouse Bcl2-Nek9 chimeric cDNA.

Fig 7

A: Using Bcl2-L1211 and Bcl2-R2i primers in PCR to amplify an RT product from Ela-mycPT1 cells resulted in not only the anticipated band of the Bcl2-β mRNA variant at the top but also a smaller band of about 500 bps (arrowhead). B: Purification, cloning and sequencing of the smaller band (indicated by the arrow in A) reveal that it is a Bcl2-Nek9 chimera (with the lowercase “t” and “a” at both ends appended by Taq DNA polymerase). The Bcl2-R2i primer (underlined region inside the sequence) has 12 nts (shaded) matched to the Nek9 sequence. There is a 5-nt unmatchable sequence (shaded 5’ACAGA3’ in boldface) after this BCl2-R2i primer. This chimera ends with another Bcl2-R2i sequence (underlined 20 nts in boldface at the 3’ end), but the Bcl2-L1211 forward primer is not in the cDNA.