Identification of differentially expressed genes in normal mucosa, adenoma and adenocarcinoma of colon by SSH

Figure 1 The results of secondary PCR amplification: Lane 1: Subtracted A-N cDNA library; Lane 2: Unsubtracted A cDNA library; Lane 3: Subtracted T-A cDNA library; Lane 4: Unsubtracted T cDNA library; Lane 5: Subtracted T-N cDNA library; Lane 6: Control; A: Subtracted A-N cDNA library; B: Subtracted A-N cDNA library; C: Subtracted A-N cDNA library.

Figure 2 Analysis of subtraction efficiency: Lanes 1-4 were performed on subtracted cDNA libraries, Lanes 5-8 were performed on unsubtracted cDNA libraries. Cycles for 1&5, 2&6, 3&7 and 4&8 were 18, 23, 28 and 33 respectively.

Figure 3 Clone PCR amplification with an average insert size of 0. 2-1 kb.

Figure 4 Characteristion of randomly selected differentially expressed clones from the three subtracted cDNA libraries screening of forward subtraction. Clone name, insert size, sequence identity and redundancy are Shown

Figure 5 Differentially screening of clones using both forward (left) and backward (right) subtracted cDNA libraries. A1: Subtracted A-N cDNA library; A2: Unsubtracted A cDNA library; B1: Subtracted T-A cDNA library; B2: Unsubtracted T cDNA library; C1: Subtracted T-N cDNA library; C2: Unsubtr acted T cDNA library.

Figure 6 Virtual Northern Blot analysis of several differentia lly expressed cDNA: N: normal mucosa; A: adenoma; T: adenocarcinoma.