Details+of+scFv+Construction+070710

Details of scFv Construction 7-7-10

Primer Sequences

// Location of Primer Order Form: smb://peptide.biodesign.asu.edu/research/CIM/Mol_Biol_Tech/Phage%20display/phage%20display%20library // // /primer%20order/Invitrogen_ Oligo Order_042209.xls //

Invitrogen_Oligo Order 042209

//Source of Primer Sequences: Optimization of primer sequences for mouse scFv repertoire display library construction //

Optimization of primer sequences for mouse scFv repertoire display library construction

//Note that the first part of the sequences do not correspond to the sequences of the primers that Tien ordered because the first part of their sequences matched with the pHEN1 vector whereas we are using the pCANTAB5 vector. The last part of the sequence matches though.//

Note that the primers actually used for the first round of scFv gene construction are a little bit different than the primers in the Invitrogen_Oligo Order 042209. Many of the primers are the same, but there were some primers that were not used that were in the 042209 form and there were also some additional primers. Primers Used for 1st Construction of scFv Genes

// Details of scFv Construction 7-7-10 // // Primer Sequences // // [[../5-27-10%20Records/Map%20of%20Phage%20Library%20Box%205-27-10.ods|Location of Tien's phage info: smb: //What are the sequences of the mouse immunoglobulin genes?10-26-10The Zhou paper constructed their primers to match the sequences in the Kabat et al database. Specifically they used the sequences from Sequences of Proteins of Immunological Interest 4th edition 1987 pg. 508-553.I have found several sources of antibody sequences.THE INTERNATIONAL IMMUNOGENETICS INFORMATION SYSTEMhttp://imgt.cines.fr/ [] NCBI IgBlast [] http://www.kabatdatabase.com/index.html I may want to look at the following paper: [|Accessing the Kabat antibody sequence database by computer] I checked out the following book from the library: Sequences of Proteins of Immunological Interest 5th edition 1991