BDB -BiopanningDataSet Details

Biopanning Data

BiopanningDataSet ID: 2558

Peptides:

Sequences
GRHDSLLR(5) ARKDALTR(1)

Sequences Features:

Motif:

Unique Sequence Number:

Experimental Method:

Phage display (subtractive panning)

Sequencing Platform:

Sequencing Method:

Sanger sequencing

Rounds of Panning:

Reference:

PMID: 10077584

Target Name:

Hairpin DNA

Template Name:

Not determined.

Structure of Target-Template Complex:

Not determined.

Structure of Target-Peptide Complex:

Not determined.

Library Name:

pComb3H X5LX2 phage display library

Affinity Measurement Method:

Affinity Measurement Description:

Brief Description:

Hairpin target oligonucleotides had the sequence 5'-Biotin-GGACGCN'N'N'CGCGGGTTTTCCCGCGNNNGCGTCC-3', where NNN was the 3-nt finger-2 target sequence and N'N'N' its complement. The selection target site is GTG. A similar nonbiotinylated oligonucleotide, in which the target sequence was TGG (compTGG), was included at 7.2 nM in every round of selection to select against contaminating parental phage. Two pools of nonbiotinylated oligonucleotides also were used as competitors: one containing all 64 possible 3-nt targets sequences (compNNN), the other containing all of the GNN target sequences except for the current selection target (compGNN). These pools typically were used as follows: round 1, no compNNN or compGNN; round 2, 7.2 nM compGNN; round 3, 10.8 nM compGNN; round 4, 1.8 mM compNNN, 25 nM compGNN; round 5, 2.7 mM compNNN, 90 nM compGNN; round 6, 2.7 mM compNNN, 250 nM compGNN; round 7, 3.6 mM compNNN, 250 nM compGNN. The sheared herring sperm DNA was first incubated with the library, before the addition of 72 nM biotinylated hairpin target oligonucleotide. Each selection was performed on streptavidin-coated magnetic beads.

BiopanningDataSet Comments:

Curator:

Curation Date:

0000-00-00

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