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Centri-Sep 96 filter plates have been developed and optimized to remove the excess dye terminators from the extension products in the Big Dye®Terminator cycle sequencing reactions. Sample volumes of 5 to 20 microliters are efficiently purified in less than 10 minutes. Purified samples result in clean electrophoretic results and long base reads. Transferring the samples to the plates is an extremely important part of the procedure and is essential to getting good quality sequencing results.

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Reduced recovery of protein samples may occur when using Centri-Spin spin columns due to specific or non-specific interaction with the column matrix. Below is a list of common modes of interaction and suggestions for minimizing their effect on sample recovery.

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Introduction:

CENTRI-SEP 96 multi-well plates are an excellent solution to the difficulties encountered in removal of excess dye labeled terminators from dye terminator reactions. The hydrated matrix in these plates has been optimized for use with Big Dye reactions. When properly used, CENTRI-SEP 96 plates will yield sequencing reactions equivalent in cleanliness to single CENTRI-SEP spin columns. Gel images obtained using reactions prepared with CENTRI-SEP 96 plates will be free of interfering smears, blobs, and gel haze caused by the presence of excess dyes. Sequences may be read from the first base and longer read lengths may be obtained due to reduced background. These improved reads result in a lower cost per base than less effective precipitation methods.

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Introduction:

CENTRI-SPIN spin columns provide a convenient and rapid method to remove excess label from nick translation or PCR labeling reactions (see Application Note A-1). For example, CENTRI-SPIN-20 will remove up to 99% of free label from a nick translation reaction in a simple 2 minute centrifugation. CENTRI-SPIN columns are not routinely tested for the presence of RNase and the majority of applications have been developed for proteins and DNA. However, the spin columns may also be used for RNA labeling reactions if precautions are taken to reduce the possibility of RNase contamination. Some of these precautions are discussed below.

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