FragmentGENE’s double-stranded linear DNA fragments offer 100% sequence fidelity and high DNA purity making them an optimal choice for a variety of downstream applications including gene assembly and other synthetic DNA needs.
To test the sequence quality, 22 different DNA fragments ranging from 300-1200bp in size with GC content between 30-70% were cloned into our in-house shuttle vector pUC57-Kan using recombination cloning and transformed into the E.coli strain Top10. Four colonies for each fragment were picked for Sanger sequencing and each clone was sequenced bi-directionally using the universal primers M13F(-47) and M13R.
FragmentGENE DNA fragments produced 92% cloning accuracy and 100% sequence fidelity. Of the 22 fragments, 14 fragments produced 100% correct clones (56 out of 56), 6 fragments had 83% correct clones (20 out of 24) and remaining 2 fragments had 65% correct clones (5 out of 8). These sequencing results confirm that FragmentGENE DNA fragments provide high cloning accuracy (>90%) and increase the efficiency of your research by minimizing the cost and allowing you to rapidly generate the DNA constructs for your downstream applications.
Figure 1: FragmentGENE gene fragments’ superior quality increases efficiency, cuts costs, and saves time by reducing downstream screening. 22 fragments ranging from 300–1200 bp with GC content between 30-70% were tested for cloning accuracy and sequence fidelity. Cloned fragments were screened using Sanger sequencing by picking four colonies for each fragment. The average cloning accuracy was >90% and 100% sequence fidelity.
|Sequence Length*||Quantity||Completion Time**||Predicted Cloning Accuracy||Sequence Fidelity|
|100-750 bp||500 ng||3-5 business days||>90%||100%
|750-1,000 bp||1,000 ng|
|1,001-2,000 bp||5-8 business days|
*Only sequences with overall GC content between 20% and 80% qualify for FragmentGENE service. GC/AT rich or sequences with complex repeats do not qualify.
**Orders containing more than 50 fragments will require additional turnaround time; Shipping time is expected to add 1-2 business days to completion time.
Standard deliverables include 500-1,000 ng of lyophilized double-stranded DNA, depending on sequence length.