AlanFung:LabNotes/CTCF/2011-6-17

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Revision as of 18:50, 17 June 2011 by >Alan6017518 (→‎Results)
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Objective

  • Perform Sanger Sequencing on library prepared by Methyl-Seq DNA Sample Prep Kit
  • Ensure the library is well constructed and that we are able to generate what Epicenter claimed to be the problem

Protocol

TA Cloning

Ligation of PCR vector

Transformation

PCR amplification and preparation for Sanger Sequencing

File:ZhangLab 2 2011-06-16 16hr 31min.jpg

  • Picked clone #1-6 for sanger sequencing

Sequencing Results

Media:06172011_ab1.zip

  • #5 failed to pass the QC due to non specific results

Results

Analysis Clone #3 for it has the highest quality score

  • Extract Sequence
NNNNNNNNNNNNAGGGCGATTGGGCCCTCTAGATGCATGCTCGAGCGGCCGCCAGTGTGATGGATATCTGCAGAATTCGGCTTCCTTGCCAGCCCGCTCAGATATAAACAAAAATATCTAAACATAATAACTCGCGCCTATAATCCCAACACTTTAAAAAACCGATTTAAACAAATCACAAAATCTAAAATTAAAAACCAACCTAACCAACATAATAAAACTTCATCTCTACTAAAAATAAAAAATTAACTGAATATAATAACAAACACCTATAATCCTAACTACTCAAAAAACTAAAAAAAAAACTGTCTCTTATACACATCTCTGATGGCGCGAGGGAGGAAGCCGAATTCCAGCACACTGGCGGCAGTTACTAGTGGATCCGAGCTCGGTACCAAGCTTGGCGTAATCATGGTCATAGCTGNTTCCTANNTACAANNCTGTNNTTAACCCCCTGGATAGGGTGAGCTGGAGTAACACTCTCCTGTTTTTTGACAGGGATGATNACGCACAGGTTGCATGGCCTACCCAGGAATTAACGCANCACCNCAAAGTTTCATTAATCCTCACCTCANGCTCTGTAAAGGAAGTTGCGAATCATTTAACCCAAGCTATTTATTTTGATTTATATGGATTTTTGGTTGCCAAATTTCCAATAAACTATTTGCCGGCGCGGNTTTTTCNNCANGGAAGCGTTTGCAAATTAATGAGGTGACTGTATTCCNCGACTTATTAAATTTGGCGGANGANAAAGANNTTCANCNNNTTAAAAAAACCCCCAAAATCAATCNCANNANTCGCCATAACNAGTNTATCNATCTCCNATAANTCAANCTNNCGNNNNNNNCAACTTACCNNNNNCTGNNNNCANCCCTNGNAANNNGNA