Use of engineered thrombin cleavage site for determination of translational reading frames.

similar yield as the standard method. Therefore, using this modified method, we can reduce the soaking time from 15 h to about 10 min. In summary, a simple modification of the crush-and-soak method allows the recovery of ODN from polyacrylamide gel within 30 min with high yield. The key difference in our procedure is the replacement of the overnight soak with a heat/freeze step (summarized in Table 2). We assume that heating causes the crushed gel to swell rapidly, hastening the diffusion of the ODN. Subsequent freezing may shrink the solid gel matrix while expanding the water volume, thus squeezing the ODN out of the gel network. This modified method is simple and fast, yet produces similar recovery as the standard overnight soaking method. It provides a rapid alternative to the standard crushand-soak method for recovering either labeled or unlabeled oligonucleotides from polyacrylamide gel.