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In this step, particular care needs to be taken to ensure that different RNAseq protocols are processed with the strand of capture in mind.
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Briefly, 1 ug of RNA was processed with the Superscript III First strand kit (Invitrogen) to obtain cDNA.
mRNAs were treated with Calf Intestinal Phosphatase (CIP) to dephosphorylate uncapped 5' ends and processed with the GeneRacer kit for first-strand synthesis of full-length cDNAs using SuperScript III reverse transcriptase (RT).
Fully processed, with the needle fans separated into individual strands.
Stretch the strand of hair.
Repeat the process with four strands of wire, etc. Decorate.
These first time voters are well-equipped to sort through and process the multiple strands of communications and propaganda.
The subsequent data were processed with Perl scripts to identify strands (i.e., sequences of three or more residues long) with two or more hot spot residues (ΔΔ G > 1.0 REU).
Meyer's breakthrough came in developing a method to start the sequencing process with single strands of DNA instead of double strands, as is usually done.
Briefly, aligned files were processed with SAMtools and subsequently VarScan (V.2.2) 13 for the detection of A to G (positive strand) or T to C (negative strand) substitutions.
This self-generated cDNA becomes amplified later in the PCR reaction and this process interferes with the accuracy of strand specificity when both strands are co-expressed.
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