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cDNA library: Zebrafish SJD day 8 fin regeneration

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   1. Zebrafish SJD day 8 fin regeneration

Zebrafish SJD day 8 fin regeneration

Name: Zebrafish SJD day 8 fin regeneration
Library ID: 1764
Organism: Danio rerio
Gender: male
Age: 0
Organ: fin
Tissue: caudal fin, regeneration after 8 days
Host: DH10B
Vector: pAMP1
Vector type: phagemid
Insert digest: UDG
Stop Codon Status: without
Description: First strand cDNA synthesis was primed using oligo-dT on magnetic beads with an additional primer 5'-gcggccgctaatacgactcacta-taggg-3'. Second strand synthesis was a 3-cycle PCR using the primers 5'-ggccgctaatacgactcactatag-3' and 5'aagcagtggtaacaacgcagagtactttttttttttttttvn-3'. cDNA was subsequently amplified in a 7-cycle PCR with the following primers: 5'-ggccgctaatacgactcactatag-3' and 5'-aagcagtggt-aacaacgcag. Deoxy-UMP adaptors were added in a third PCR (5 cycles) and the primers 5'-caucaucaucauggccgctaatacgactcactataggg-3' and 5'-cuacuacuacuaaagcagtggtaacaacgcagagtac-3'. Ends were treated with uracil DNA glycosylase and product with 3' overhangs was annealed to complementary ends of pAMP1. Insert can be excised using EcoRI and NotI. Library constructed by Joe Barnes and Steve Johnson (Washington University).

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