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Hello all,
I'm just starting my thesis work and have been looking for ways preparing RNA-seq libraries (specifically for Illumina). As I work on yeast I have the option to multiplex quite heavily and have reached a point where library preps will cost more than the sequencing itself. Thus, I've been looking for cheap and high throughput ways to get to decent libraries.
I ran across Miltenyi Biotec's cDNA synthesis kit and it looks like it would be a great starting point, but haven't found a lot of information on the quality of the cDNA produced. I know their process is first strand synthesis only, but that is easy enough to fix/not a problem for some library prep procedures. I was wondering if anyone had experience with this kit and if so, how did they go about preparing libraries afterwards. Also, does anyone know of a relatively cheap method for preparing libraries in bulk? I've managed to get down to ~$45 using NebNext kits ordered through my university, but it is a bit daunting to think of the number of gels I'll have to go through.
Thanks all,
Metz
I'm just starting my thesis work and have been looking for ways preparing RNA-seq libraries (specifically for Illumina). As I work on yeast I have the option to multiplex quite heavily and have reached a point where library preps will cost more than the sequencing itself. Thus, I've been looking for cheap and high throughput ways to get to decent libraries.
I ran across Miltenyi Biotec's cDNA synthesis kit and it looks like it would be a great starting point, but haven't found a lot of information on the quality of the cDNA produced. I know their process is first strand synthesis only, but that is easy enough to fix/not a problem for some library prep procedures. I was wondering if anyone had experience with this kit and if so, how did they go about preparing libraries afterwards. Also, does anyone know of a relatively cheap method for preparing libraries in bulk? I've managed to get down to ~$45 using NebNext kits ordered through my university, but it is a bit daunting to think of the number of gels I'll have to go through.
Thanks all,
Metz