Figure 3

Religation at low concentration lead to non-proportional amplification. (a) Re-digestions of HpaII digested, religated and amplified samples. Religations were done at different concentrations. The concentrations go down in descending order from number 1 to 12. Six different concentrations were chosen i.e. 12 ng/μ L, 10 ng/μ L, 8 ng/μ L, 4 ng/μ L, 1 ng/μ L, 100 pg/μ L. Odd numbers represent the amplified samples redigested with the same enzyme (HpaII) while even numbers represent same samples redigested with a different enzyme (AflIII). (1,2: 12 ng/μ L, 3,4: 10 ng/μ L, 5,6: 8 ng/μ L, 7,8: 4 ng/μ L, 9,10: 1 ng/μ L, 11,12: 100 pg/μ L). The asterisks represent the fragments that are over amplified as compared to others. 10% polyacrylamide gels stained with EB were used for redigested samples. (b) Quantitative scanning of relative band intensities. The arrows shown in the scan of lane-11, corresponding to the asterisks in fig. 3a, represent DNA fragments that are relatively better amplified as compared to others in the same lane.