1-23-13+Sequencing+results

1-23-13 Sequencing Results

Received sequencing results after I attempted to linearize the pcomb plasmid and perform an in-fusion with a pca constructed fragment. The purpose of this was to try to readd the stop codon that had been deleted during previous modifications (see 11-5-12 Correct deletion of stop codon)

Sequencing results found here "C:\kurt\storage\CIM Research Folder\DR\2013\1-23-13\pcomb fusion sequencing check 1-23-13"

Three colonies were sent for sequencing from each group. The groups were as follows.

1 previous ethanol precipitated vector in-fusion bottom band (gel of bands on 1-14-13 of notebook) 2 previous ethanol precipitated vector in-fusion middle band (gel of bands on 1-14-13 of notebook) 3 previous ethanol precipitated vector in-fusion top band (gel of bands on 1-14-13 of notebook) 4 vector in-fusion bottom band (gel of bands on 1-16-13 of notebook) 5 vector in-fusion middle band (gel of bands on 1-16-13 of notebook) 6 vector in-fusion top band (gel of bands on 1-16-13 of notebook)

Which plasmids had the correct sequencing result? 1p1 (this sequence does not seem to be quite right; it has 1 bp deleted at 2628 of what pcomb3xssv3 should be) 1p2 1p3 4p1 4p2 4p3 5p1 6p2

region of interest TTAATTTAAGGCCGCAGATCTGGGAAATTGTAAGCGTTAATATTTTG

As soon as I receive the Primers to sequence whole pcomb3XSSv3 then I can choose a few of these plasmids to verify the whole sequence.

2-7-13

Sequencing results for whole plasmid C:\kurt\storage\CIM Research Folder\DR\2013\2-7-13\pcomb_sequences

Original Sequencing files "C:\kurt\storage\CIM Research Folder\DR\2013\2-7-13\pcomb_sequences\original sequencing files"

2-12-13 I handed this off to Mara and she started working on it. Her files can be found here S:\Research\Cancer_Eradication\Discovering tumor specific antigens\Phage Library\2-12-13 pcomb sequences