Figure 1

The analysis on the SSH library. Analysis of ds cDNA synthesis products and Rsa I digestion. Cotton double-stranded cDNA before (Lane 2) and after (Lane 1) Rsa I digestion. Lane M: DNA size markers. B) Analysis of ligation efficiency. Lane 1: PCR products using Tester 1-1 (Adaptor 1-ligated) as the template and the Histone 3 3' primer and PCR primer 1. Lane 2: PCR products using Tester 1-1 (Adaptor 1-ligated) as the template, and the Histone 3 3' and 5' primers. Lane 3: PCR products using Tester 1–2 (Adaptor 2R-ligated) as the template, and the Histone 3 3' primer and PCR primer 1. Lane 4: PCR products using Tester 1–2 (Adaptor 2R-ligated) as the template, and the Histone 3 3' and 5' primers. Lane M: DNA size markers. C) Results of PCR-select cDNA subtraction analysis. Lane 1: Forward secondary PCR products of unsubtracted. Lane 2: Forward secondary PCR products of subtracted. Lane 3: Reverse secondary PCR products of unsubtracted. Lane 4: Reverse secondary PCR products of subtracted. Lane M: DNA size markers. D) Analysis of subtraction efficiency using PCR. The subtracted and unsubtracted pools of cDNA were amplified by using primers for the constitutively expressed Histone 3 gene. PCR was performed on the subtracted (Lanes 1–4) or unsubtracted (Lanes 5–8) secondary PCR product with the Histone 3 5' and 3' primers. Lanes 1 & 5: 18 cycles; Lanes 2 & 6: 23 cycles; Lanes 3 & 7: 28 cycles; Lanes 4 & 8: 33 cycles.