Hey all,
I've been using the Nextera XT kit without bead normalization and have been getting inconsistent yields after the PCR step. There doesn't seem to be any rhyme or reason to it. Right now, I'm trying to get some decent libraries with plasmids. Did 5 preps and only two of them were above 2nM. I've had the same issue with bacterial genomes and commercially prepared human genome. We've had limited success increasing input to 6ng and using 90ul Ampure beads but that too is inconsistent.
Has anybody had the same problem and if so, what have you done to fix it? I've been advised by another lab to switch to the regular Nextera kit but money is an issue with our lab. If we can make the cheaper Nextera XT kit work consistently, it would be better. Any advice would be appreciated!
BigAlPal
I've been using the Nextera XT kit without bead normalization and have been getting inconsistent yields after the PCR step. There doesn't seem to be any rhyme or reason to it. Right now, I'm trying to get some decent libraries with plasmids. Did 5 preps and only two of them were above 2nM. I've had the same issue with bacterial genomes and commercially prepared human genome. We've had limited success increasing input to 6ng and using 90ul Ampure beads but that too is inconsistent.
Has anybody had the same problem and if so, what have you done to fix it? I've been advised by another lab to switch to the regular Nextera kit but money is an issue with our lab. If we can make the cheaper Nextera XT kit work consistently, it would be better. Any advice would be appreciated!
BigAlPal
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