Methods of Amplifying Paired Transcript Sequences from Single Cells
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United States of America Patent
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Issued Date -
Mar 16, 2023
app pub date -
Sep 12, 2022
filing date -
Mar 14, 2020
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Abstract
The disclosure provides methods for amplification of paired transcript sequences from a single cell, such as, but not limited to alpha and beta T cell receptor (TCR) sequences from a single T cell. The method can include: placing a single T cell into a cell lysis solution to provide a T cell lysate comprising RNA; generating first strand cDNA from the RNA; amplifying the first strand cDNA to provide amplified cDNA; and amplifying the alpha and beta TCR sequences from the amplified cDNA in a single reaction, wherein the alpha and beta TCR sequences are amplified using three sets of TCR amplification primers. In some instances, the methods can be performed using no more than six sets of primers.
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