Methods for preparing a sample for nucleic acid amplification using tagmentation
AssigneeIllumina Cambridge Limited
InventorsLouise Fraser, Paula Kokko-Gonzales, Andrew Slatter
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Abstract
Presented are methods and compositions for preparing samples for amplification and sequencing. Particular embodiments relate to methods of preparing nucleic acid-continuing cellular samples for library amplification, wherein the methods include lysing cells of the sample to form a lysate, amplifying the nucleic acids from the lysed samples, exposing the amplified nucleic acids to a solid surface, and clonally amplifying the amplified nucleic acids to generate clusters.
Claims25
A method of preparing a sample for library amplification and subsequent amplification comprising the following steps:
- on a provided nucleic acid-containing cellular sample,
- (a) lysing cells of the sample with a lysis reagent to liberate nucleic acid from within the cells of the cellular sample, thereby forming a lysate;
- (b) amplifying the nucleic acids from the lysate to form amplified nucleic acids;
- (c) exposing the amplified nucleic acids to a solid surface with immobilized amplification primers, thereby immobilizing the amplified nucleic acids on the solid surface;
- and
- (d) clonally amplifying the immobilized, amplified nucleic acids on the solid surface to generate clusters;
- wherein there is no purification of the nucleic acid from the lysate prior to beginning the amplification step (b);
- and
- wherein amplifying the nucleic acid in step (b) comprises tagmentation,
- wherein the sample is a blood sample, a whole blood sample, or a dried blood sample.
A method of preparing a sample for library amplification and subsequent amplification comprising the following steps:
- on a provided nucleic acid-containing cellular sample,
- (a) lysing cells of the sample with a lysis reagent to liberate nucleic acid from within the cells of the cellular sample, thereby forming a lysate;
- (b) amplifying the nucleic acids from the lysate to form amplified nucleic acids;
- (c) exposing the amplified nucleic acids to a solid surface with immobilized amplification primers, thereby immobilizing the amplified nucleic acids on the solid surface;
- and
- (d) clonally amplifying the immobilized, amplified nucleic acids on the solid surface to generate clusters;
- wherein there is no purification of the nucleic acid from the lysate prior to beginning the amplification step (b);
- and
- wherein amplifying the nucleic acid in step (b) comprises tagmentation,
- wherein the sample is a formalin fixed paraffin embedded (FFPE) sample.
Dependencies are inferred from the claim text.