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. 2005 Jul 5;389(Pt 2):403–412. doi: 10.1042/BJ20042032

Figure 4. RT-PCR analysis of SP-B minigene mRNA in CHO cells.

Figure 4

Total RNA (1 μg) from transfected CHO cells was reverse transcribed by M-MLV reverse transcriptase, with a poly(T) primer. The reverse transcription product (1 μl) was used as a template for PCR amplification. (A) PCR amplification of different regions of SP-B mRNA from cells transfected with pBi4normal. The PCR primer pair for each lane is given above the gel. (B) PCR amplification of SP-B mRNA from cells transfected with recombinant plasmid DNAs: lane 1, pBi4del193; lane 2, pBi4del264; lane 3, pBi4del211; lane 4, pBi4normal; lane 9, pBi4del340; lane 11, vector pcDNA3.1 (control); lane 12, no plasmid DNA; lane M, 1 kb DNA ladder; lane Ct, control for PCR with distilled water. The PCR primers are given above the gel. (C) Sequence analysis of RT-PCR products amplified with primers 70A and 1194 from pBi4del193. The PCR products were purified and sequenced with primer 70A. DNA sequencing was done with an Applied Biosystems ABI 377 DNA sequencer. The junctions of exons 2 and 3, exons 3 and 4, and exon 4 and intron 4, are indicated by arrows.