Your Q&A from the GeneArt™ DNA Libraries webinar
Your colleagues submitted interesting questions during the webinar, “GeneArt™ DNA Libraries: protein engineering for every lab and budget.” We’ve collected and answered them here.
Q-Does Thermo Fisher Scientific™ also offer cloning of gene fragment libraries into customer-provided plasmids?
A- Most of our library products can be cloned and delivered in customer-provided vectors. The two exceptions are GeneArt™ Strings™ DNA Libraries, which are only delivered as linear DNA fragments, and libraries that have a biosafety level of 2 or above. In rare cases, customer-provided vectors have low transformation efficiency and are thus unsuitable for cloning libraries.
Q- Are GeneArt™ Strings DNA Libraries available with TRIM oligos?
A- GeneArt Strings DNA Libraries are only available with IUPAC- defined ambiguous DNA nucleotides. GeneArt™ Combinatorial Libraries are available using TRIM (trinucleotide mutagenesis) technologies.
A- Like the GeneArt Strings DNA Fragments, GeneArt Strings DNA Libraries are only available as uncloned, double-stranded linear DNA strands. The GeneArt Strings DNA Libraries represent pools of GeneArt Strings DNA Fragments of 200–2,000 bp, containing up to 3 blocks of degenerate nucleotides with randomized distribution.
Q- Can TOPO™ cloning technology be used with GeneArt Strings DNA Libraries?
A- Yes. GeneArt Strings DNA Libraries can be cloned with any cloning technology of choice. It is important to have the cloning method of choice pre-determined so that you order the correct type of DNA ends.
Q- Are GeneArt Strings DNA Libraries available with non-standard nucleotide mixes?
A- GeneArt Strings DNA Libraries are only available with IUPAC-defined ambiguous DNA nucleotides, which are equimolar mixes of the respective nucleotides.
Q- Can GeneArt Strings DNA Libraries be made with UTP (U or uracil) nucleotides?
A- No, only DNA bases can be incorporated into GeneArt Strings libraries.
Q- What is the maximum diversity for GeneArt Combinatorial Libraries?
A- The standard diversity that can be obtained in a GeneArt Combinatorial Library is 1011. This is defined by the amount of non-amplified library we deliver to the customer, which is calculated to be 170 fmol of DNA, or 1011 molecules. Upon customer request we can also deliver diversities of up to 1012 as an additional service.
Q- What are the design restrictions for GeneArt Combinatorial Libraries?
A- In general, there are very few design restrictions. We will consider any customer request, and in the vast majority of cases we find a solution to produce the library according to customer design. In our experience, there are some designs that make library synthesis very difficult, for example, those with highly repetitive sequences.
Q- Can GeneArt Strings DNA Libraries be optimized?
A- Yes, sequence optimization is available for all of our DNA library products.
Q- Do GeneArt DNA Strings Libraries have the same price and turnaround time as GeneArt DNA Strings Fragments?
A- No, each randomized nucleotide request incurs an extra charge. The turnaround time to produce GeneArt DNA Strings Libraries with randomized nucleotides is 10–15 business days. The turnaround time to produce GeneArt DNA Strings Fragments is 5–-8 business days, depending on the size range for the fragments.