Phage Display

You can perform 8 PCR reactions with each primer pair.
The Mouse IgG Library Primer Set contains 50 primers in 2 primer sets. The primers are ready-to-use for common PCR (conc. 10 pmol/µl in 10 mM Tris-HCl pH 8, 1 mM sodium EDTA).
With the first set of primers 25 separate reactions should be performed. Each variable heavy chain forward primer (1A-L) has to be combined with the constant region IgG reverse primer (1M). Analogous, each κ- and λ-light chain forward primer (1N-W, 1Y) has to be combined with the corresponding constant region reverse primer (1X or 1Z), respectively.
With the second primer set 25 separate reactions should be performed to add the restriction sites for cloning into the pSEX81 vector.

After reconstitution, the hyperphage can be kept at 2-8°C for several days, however, the infection potential will decrease gradually.
Hyperphage stored at 2-8°C for several weeks (up to 2 months) were still active, however, the pfu was more than 50% decreased.
For long term storage it is possible to add 20% glycerol and store the hyperphage at -80°C. This will also lead to a decrease in activity.

The concentration of our hyperphage is lot dependent. We try to keep the concentration > 1E+12 particles/ml. The specific concentration is indicated on the product label. Please contact us to ask for the concentration of your specific lot or the current lot in stock.
10 mM Tris-HCl (pH 7.5), 20 mM NaCl, 2 mM Na-EDTA
Please be informed that the packaging cell line has been patented (i.e. it is proprietary) and is unfortunately not available publicly.
Therefore it is not possible to amplify hyperphage in your lab.

TG1 is compatible with hyperphage and the classical strain for phage display. It is very suitable for panning (one panning round per day).
For the last panning round XL1-Blue MRF’ may be used, since the plasmid stability, quality and yield is higher with this strain.
Alternatively to TG1 or XL1-Blue MRF’, ER2783 is also suitable for library cloning and packaging.

It is important to use a bacterial strain with lacIq genotype (for example XL1 Blue or TOP10F’) with our pSEX81 plasmid. Bacterial strains with lacIq genotype carry the lacIq repressor that represses expression from the lac promoter and so allows cloning of toxic genes. The PIII fusion protein expressed by our pSEX81 vector is toxic to the bacterial cells.
DH5alpha or TOP10 (without F episome) are lacIq negative and therefore the toxic proteins are expressed in these cells. When using lacIq negative cell lines with the pSEX81 vector you will receive only a few colonies and most often the plasmid is somehow mutated in these colonies.
We also recommend to include 100 mM Glucose to the culture medium to avoid possible mutations of the plasmid.

Only vectors coding for a fully functional PIII protein are suitable for the hyperphage system, for example pSEX81, pComb3/pComb3X or similar vectors.
According to the literature pCANTAB vector is compatible with hyperphage, although with lower efficiency as for example pSEX81 (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1866265/).