Generating templates for sequencing

Random insertion of primer binding sites into target DNA for sequencing

Mini-Mu transposons can be used for generating templates for sequencing. In the reaction, the mini-Mu transposon is integrated in vitro into the target DNA to be sequenced. The reaction requires only one protein component, the MuA transposase, in addition to linear mini-Mu transposon DNA. The transposition reaction is highly efficient and produces relatively even distribution of integrations into the target DNA.

Applications

Efficient and easy creation of Sanger sequencing templates

Benefits

Simple: no need for primer-walking or subcloning
Efficient: generate thousands of sequencing templates from a single reaction
Universal primers: a single set of primers can be used to sequence any clone, no need for custom primers

Figure 1 (click to enlarge) Method for generating templates for sequencing using mini-Mu Transposon. Transposon (Tn) contains an antibiotic resistance (Ab^R) selection marker). From the selected clones, sequencing is carried out bi-directionally using primers specific for the ends of the transposon.

Below you can find recommended products for generating templates for sequencing

Transposase	Transposon
MuA Transposase	Cat-Mu Transposon (CamR)	Select inserts on chloramphenicol plate
MuA Transposase	Kan-Mu Transposon (KanR)	Select inserts on kanamycin plate

References

Haapa S, Suomalainen S, Eerikäinen S, Airaksinen M, Paulin L, Savilahti H (1999) An efficient DNA sequencing strategy based on the bacteriophage Mu in vitro DNA transposition reaction. Genome Res 9:308-315

Generating templates for sequencing

Random insertion of primer binding sites into target DNA for sequencing

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