Optimized conditions for cycle sequencing of PCR products.

Rapid sequencing of unpurified PCR products by thermal asymmetric PCR cycle sequencing using unlabeled sequencing primers.

Numerous methods have been developed for direct sequencing of single or double-stranded DNA amplified by PCR, and most require template purification steps prior to sequencing to remove excess unincorporated primers and dNTPs. Purification methods can be cumbersome, laborious and time consuming, or yield low DNA recoveries. Strategies for direct sequencing of doublestranded DNA products without ...

Preparation of PCR products for DNA sequencing.

We demonstrate that routine PCR product analytical agarose gels can also serve as preparative gels for quick DNA template purification before sequencing. The band of interest is excised, placed into a Gel Nebulizer inside a Micropure separator and rapidly purified in a single centrifugation step. Gel-purified PCR product, suitable for manual and automated sequencing, is delivered within 10 min.

Optimized conditions for cloning PCR products into an XcmI T-vector.

products. BioTechniques 12:28-30. 10.Scharf, S.J., G.T. Horn and H.A. Erlich. 1986. Direct cloning and sequence analysis of enzymatically amplified genomic sequences. Science 233:1076-1078. 11.Shuldiner, A.R., L.A. Scott and J. Roth. 1990. PCR-induced (ligase free) subcloning: a rapid reliable method to subclone polymerase chain reaction (PCR) product. Nucleic Acids Res. 18:1920. 12.Starr, L. a...

5'-tailed sequencing primers improve sequencing quality of PCR products.

An efficient method for purification of PCR products for sequencing.

A high-throughput DNA sequencing method that generated high quality data was developed. A frame fashioned from a standard agarose gel combined with 0.1%-0.2% low-melting point (LMP) agarose gel was used to isolate the PCR product of interest. Collected PCR products were centrifuged without any reagents and the supernatants were directly used for a sequencing reaction. This method is simple and ...