Single Pass and End Reads Sequencing

When sequencing requires any primers that are considered common. we provide them free of charge. These primers are M13F, M13R, T7 promoter, T7 terminator, and SP6. However, we caution our users to be very specific when asking for the common primers. For example, there are at least two primers that are called M13F and at least four primers that are called SP6. Please refer to sequences of common primers found in the Common Sequencing Primers submenu. For PCR fragments, customers can submit their own primers, often the same as those used to generate PCR fragments or we can synthesize them providing that the customer shares the primer information. In addition, we have primer information on about 500 different commonly used vectors, assuming that the fragment of interest is cloned into Multiple Cloning Site (MCS). You can find the information about vectors in our database under the Vector List submenu. Please be advised that sometimes the names of vectors in this submenu can be slightly different from those found in other sources. If you have any questions, please contact us.

Sequencing by Primer Walking: Construct Sequence Confirmation

If the length of a cloned insert, PCR, or any other fragment to be sequenced is longer than a typical good quality read produced by an ABI instrument (750-950 bases), you may need to add more reads to cover the entire template of interest. Additional primers will need to be submitted or designed by Wyzer Biosciences, synthesized, and reactions performed to completely sequence the entire region of interest. If not specified by a requestor, we assume that single-stranded coverage is sufficient. But in most cases, double stranded coverage will be provided. This will be noted in our report.

Publication Quality Sequencing: Double-Stranded Coverage

Sometimes Publication Quality Sequencing, double-stranded coverage over the entire region of interest, is required when submitting a paper for publication, grant application, or any other purpose. In this case, we will independently carry out primer walking on both DNA strands starting with the initial sequence information obtained from vector or customer provided specific primers. The Watson-Crick complementarity between the two DNA strands assures sufficient accuracy for Publication Quality Sequencing. If necessary, additional reads can be added to increase the accuracy of the data.

Setting up Sequencing Reaction Yourself

On occasion, a customer may elect to perform sequencing and cleanup of reactions in his/her laboratory and then ask us to run them on our instruments without additional intervention. Unless a catastrophic failure on our side occurs, we will run those reactions only once, regardless of the outcome. We will contact you and discuss additional options.