Hello Everyone,
Been a lurker for a while, but I had a question regarding some Illumina sRNA libraries I've been making and know this is the go-to-place for things of that sort. I'll make sure to make an introduction post shortly.
I'm working through the Illumina Tru-Seq sRNA protocol. I've had success in the past with the v1.5 and earlier edition of the Tru-Seq kit; but I've noticed by gels look a little different with this version (the protocol revision is May '12) of the Tru-Seq protcol than last time.
Attached are a few gel images. My input was total RNA, not purified miRNA.
Gel image 1 is of libraries I made today. Notice the very intense, thick band below the band of interest (in between the two lower bands on the three band ladder (Custom ladder). Gel Image 2 is of libraries I made a year ago using the Tru-Seq kit, albeit a different version than the one they are supplying these days. No intense band below...are those adapter dimers/concatamers?
I'm fairly certain the protocol worked as the expected band is where it should be, but I'm a little concerned by the unexpected bright signal below it. Any input would be greatly appreciated!
Been a lurker for a while, but I had a question regarding some Illumina sRNA libraries I've been making and know this is the go-to-place for things of that sort. I'll make sure to make an introduction post shortly.
I'm working through the Illumina Tru-Seq sRNA protocol. I've had success in the past with the v1.5 and earlier edition of the Tru-Seq kit; but I've noticed by gels look a little different with this version (the protocol revision is May '12) of the Tru-Seq protcol than last time.
Attached are a few gel images. My input was total RNA, not purified miRNA.
Gel image 1 is of libraries I made today. Notice the very intense, thick band below the band of interest (in between the two lower bands on the three band ladder (Custom ladder). Gel Image 2 is of libraries I made a year ago using the Tru-Seq kit, albeit a different version than the one they are supplying these days. No intense band below...are those adapter dimers/concatamers?
I'm fairly certain the protocol worked as the expected band is where it should be, but I'm a little concerned by the unexpected bright signal below it. Any input would be greatly appreciated!