Selected article for: "amplified insert and sequencing analysis"
Author: Rashmi Mohanty; Xinquan Liu; Jae You Kim; Xiujuan Peng; Sahil Bhandari; Jasmim Leal; Dhivya Arasappan; Dennis C. Wylie; Tony Dong; Debadyuti Ghosh
Title: Identification of peptide coatings that enhance diffusive transport of nanoparticles through the tumor microenvironment
  • Document date: 2019_6_4
    • ID: e2uzk4u1_73
    Snippet: Briefly, the random insert region of the T7 library DNA was amplified from the denatured phage sample by PCR using 2× KAPA HiFi HotStart ReadyMix (KAPA Biosystems). An additional PCR was performed to attach dual indices and overhanging adapters that would be used to recognize individual samples from the final pooled library of all samples (i.e. each round from each replicate pooled together). The primers for the barcoding were provided from the .....
    Document: Briefly, the random insert region of the T7 library DNA was amplified from the denatured phage sample by PCR using 2× KAPA HiFi HotStart ReadyMix (KAPA Biosystems). An additional PCR was performed to attach dual indices and overhanging adapters that would be used to recognize individual samples from the final pooled library of all samples (i.e. each round from each replicate pooled together). The primers for the barcoding were provided from the Nextra XT Index kit (FC-131-1001, Illumina). The PCR amplicon was purified from the free primers using AMPure XP beads (Beckman Coulter) after each PCR amplification step. The size of the purified PCR product was verified by 2% agarose gel electrophoresis, and the DNA concentration was quantified using a Nanodrop spectrophotometer (Thermo Fisher Scientific) after each PCR step. After purification, each sample was diluted to 40 nM with 10 mM Tris buffer, pH 8.5. One microliter of each sample was pooled together, and the library was run on Illumina MiSeq (The University of Texas at Austin (UT-Austin) Genomic Sequencing and Analysis Facility).

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