COMPOSITIONS AND METHODS FOR END TO END CAPTURE OF MESSENGER RNAS

The subject matter disclosed herein is generally directed to methods and compositions for a single- or multi-pot protocol for the efficient end to end capture of RNAs (inclusive of their poly- A tail or their 3' end). The invention includes the use of capture oligonucleotides containing a 3...

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Bibliographische Detailangaben
Hauptverfasser:	ZWIRKO, Zachary, AL'KHAFAJI, Aziz, HACOHEN, Nir
Format:	Patent
Sprache:	eng ; fre
Schlagworte:	BEER BIOCHEMISTRY CHEMISTRY COMPOSITIONS OR TEST PAPERS THEREFOR COMPOSITIONS THEREOF CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES CULTURE MEDIA ENZYMOLOGY MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS METALLURGY MICROBIOLOGY MICROORGANISMS OR ENZYMES MUTATION OR GENETIC ENGINEERING PROCESSES OF PREPARING SUCH COMPOSITIONS PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS SPIRITS VINEGAR WINE
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Beschreibung
Zusammenfassung:	The subject matter disclosed herein is generally directed to methods and compositions for a single- or multi-pot protocol for the efficient end to end capture of RNAs (inclusive of their poly- A tail or their 3' end). The invention includes the use of capture oligonucleotides containing a 3' non-extendable end and a selectively cleavable base upstream of an oligo-dT or oligo-dN and a 5' sequence containing unique molecular identifiers, and 2) a deoxyuracil glycosylase that acts only on a deoxyuracil present in a DNA:DNA duplex or DNA/RNA heteroduplex. The invention also includes the use of a dual template switching mechanism.. L'objet de l'invention concerne de manière générale des méthodes et des compositions pour un protocole monotope ou multipot en vue de la capture d'ARN de bout en bout efficace (y compris leur queue poly-A ou leur extrémité 3'). L'invention concerne également l'utilisation d'oligonucléotides de capture contenant une extrémité non extensible 3' et une base sélectivement clivable en amont d'un oligo-dT ou d'un oligo-dN et d'une séquence 5' contenant des identifiants moléculaires uniques, et 2) une désoxyuracile glycosylase qui agit uniquement sur un désoxyuracil présent dans un duplex ADN:ADN ou un hétéroduplex ADN/ARN. L'invention concerne en outre l'utilisation d'un mécanisme de commutation à double modèle.