Details of scFv Construction 070710

2015-01-13

azim58 - 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:
[[../5-25-10%20Records/Phage%20Library%20Planning/Optimization%20of%20prime
r%20sequences%20for%20mouse%20scFv%20repertoire%20display%20library%20const
ruction.pdf|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|Locat
ion 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/http://www.bioinf.org.uk/abs/
NCBI IgBlast
http://www.ncbi.nlm.nih.gov/igblast/showGermline.cgi
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

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