DePIE

Design Primers for Protein Interaction Experiments

Step 1. Select ONE Input Option Using the Following Fields

1. Upload a file with NCBI protein accession numbers from your local computer:

(Click here to see input file format)

2. OR enter NCBI protein accession numbers manually:

(One accession number per line)

Step 2. Add Restriction Sequences to the Primers (Optional)

Protein interaction experiments usually require the cloning of CPR products into a palmed for expression. It is thus necessary to have an option to input restriction sequences, which are added priori to the 5'-end of primer segments. By default the program uses the GATEWAYTM clones restriction sequences. The defaults can be changed when necessary. To do so, change sequences in the following text areas to those that you want.

1. 5'-end restriction sequence:
   

2. 3'-end restriction sequence:
   

1. G/C content (35% to 65% as default)

2. Annealing temperature (45°C to 75°C as default)

3. G/C clamps Yes No

4. At the 3'-end, no more than 2 3 4 G or C bases

5. Secondary structure (the number of contiguous base pairs of homology to itself or to its counterpart)

6. Mispriming (the number of contiguous base pairs of homology to its target sequence)

1. Nucleotide sequences?   Yes No

2. Amino acid sequences?   Yes No

3. PSORT predictions (signal peptide, transmembrane segment, topology)?   Yes No