US2020181602A1PendingUtilityA1

Assay methods and compositions for detecting contamination of nucleic acid identifiers

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Assignee: AGILENT TECHNOLOGIES INCPriority: Jul 10, 2017Filed: Feb 17, 2020Published: Jun 11, 2020
Est. expiryJul 10, 2037(~11 yrs left)· nominal 20-yr term from priority
Inventors:Katie Leigh ZobeckPaige AndersonJavelin ChiHenrik Johansson
A63F 13/55C12Q 1/6848C12Q 1/6806C12Q 1/689C12Q 1/6874C12N 15/1065
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Claims

Abstract

The present invention relates to nucleic acid samples for massively parallel sequencing. More particularly, the present invention relates to assay methods, compositions and kits for detecting contamination of nucleic acid identifiers such as sample barcodes.

Claims

exact text as granted — not AI-modified
1 .- 18 . (canceled) 
     
     
         19 . A composition comprising:
 at least one oligonucleotide having a 5′ constant region, a sample identifier, and a 3′ constant region, and   at least one assay primer comprising a priming portion and an assay identifier.   
     
     
         20 . The composition of  claim 19 , further comprising one or more of:
 a DNA polymerase, and   deoxynucleotides.   
     
     
         21 . The composition of  claim 19 , wherein the 5′ constant region comprises a standard 5′ amplification region for a sequencing platform and a sequencing priming region. 
     
     
         22 . The composition of  claim 21 , wherein the standard 5′ amplification region comprises a P5 sequence. 
     
     
         23 . The composition of  claim 21 , wherein the standard 5′ amplification region comprises a P7 sequence. 
     
     
         24 . The composition of  claim 19 , wherein the 3′ constant region comprises a standard 3′ amplification region for a sequencing platform. 
     
     
         25 . The composition of  claim 24 , wherein the standard 3′ amplification region comprises a P5 sequence. 
     
     
         26 . The composition of  claim 24 , wherein the standard 3′ amplification region comprises a P7 sequence. 
     
     
         27 . The composition of  claim 19 , further comprising a second primer. 
     
     
         28 . The composition of  claim 27 , wherein the second primer comprises a 3′ region the same or complementary to the 5′ constant region of the at least one oligonucleotide. 
     
     
         29 . The composition of  claim 28 , wherein the second primer further comprises a 5′ region comprising a standard amplification region for a sequencing platform. 
     
     
         30 . The composition of  claim 29 , wherein the 5′ region of the second primer allows for sequencing on a sequencing platform not supported by the sequence of the 5′ constant region of the at least one oligonucleotide. 
     
     
         31 . The composition of  claim 19 , wherein the oligonucleotide is a library molecule. 
     
     
         32 . The composition of  claim 19 , comprising at least 8 oligonucleotides. 
     
     
         33 . The composition of  claim 19 , wherein each of the at least one oligonucleotides has the same sample identifier, unless the composition is contaminated. 
     
     
         34 . The composition of  claim 19 , wherein the sample identifier is a sample barcode. 
     
     
         35 . The composition of  claim 34 , wherein the sample barcode comprises 6 or more random or degenerate nucleotides. 
     
     
         36 . The composition of  claim 34 , wherein sample barcode comprises 4, or more, known nucleotides. 
     
     
         37 . The composition of  claim 19 , wherein the assay identifier comprises 10 random or degenerate nucleotides.

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