? TROUBLESHOOTING 87Estimation of the cleavage intensity. Measure the integrated intensity of the PCR amplicon and cleaved bands by using ImageLab, ImageJ or other gel quantification software.88For each lane, calculate the fraction of the PCR product cleaved (fcut) by using the following formula: fcut = (b + c)/(a + b + c), where a is the integrated intensity of the undigested PCR product and b and c are the integrated intensities of each cleavage product. A sample gel is shown in Figure 6.89Indel occurrence can be estimated with the following formula, based on the binomial probability distribution of duplex formation: indel(%)=100×(1−(1−fcut))
