SIMPLIFIED SAMPLE PREPARATION FOR RNA ANALYSIS
First Claim
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1. A method for selective priming during reverse transcription of RNA to cDNA, comprising contacting an RNA sample with one or more primer oligonucleotides, wherein said primer oligonucleotides are comprised of one or more oligonucleotide sequences which hybridize specifically to mRNA.
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Abstract
Methods and kits for selective preparing cDNA relatively free of sequences found in rRNA and subcellular RNAs are disclosed. The methods and kits utilize approximately 200 hexamer sequences which target messenger RNA. The methods and kits are useful in preparing samples for sequencing analysis, especially when performing single molecule sequencing by synthesis.
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19 Claims
- 1. A method for selective priming during reverse transcription of RNA to cDNA, comprising contacting an RNA sample with one or more primer oligonucleotides, wherein said primer oligonucleotides are comprised of one or more oligonucleotide sequences which hybridize specifically to mRNA.
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17. A kit for selective priming during reverse transcription of RNA to cDNA, comprising one or more primer oligonucleotides, wherein said primer oligonucleotides are comprised of one or more oligonucleotide sequences which hybridize specifically to mRNA.
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18. A pre-selected mixture of random hexamer sequences, comprising a plurality of hexamer sequences comprising those sequences containing no greater than 4 C+G per hexamer;
- or a plurality of hexamer sequences provided in Table 1 when the sample is of human origin.
- View Dependent Claims (19)
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