ABI releases revised spin column protocol for difficult samples when using Big Dye v. 3.0.
Introduction:
Users of Big Dye version 3 chemistries have reported occasional dye blobs on sequence images even after cleanup of reactions using spin columns or plates. The blobs are apparently due to aggregation of the dyes. ABI has released a modification of the cleanup protocol supplied with the version 3 chemistry. The recommendation is to add 1/10th volume of 2.2% SDS to the sample, mix, heat to 95 degrees C for 5 minutes, cool to ambient temperature and spin briefly to collect sample in bottom of tube.. The sample is then added to the spin column as previously described. This additional step is also recommended for samples processed using Centri·Sep 96 plates.