Hello Everyone,
This is my first post in this forum. Hooray!! Anyways, on to my question:
My lab is just beginning to get into RNA sequencing and we are currently working on our library preparation technique for sequencing using the illumina platform (GA2). In my travels through the literature on this topic I have noticed that several papers have suggested that during the post-amplification size selection of the library preparation, the band size should be relatively narrow. In other words if you are sequencing with 300 bp fragments it is better to choose a range of 300bp +/-50bp as opposed to +/-100 bp. Why is this? Is it okay to size select for the full range of fragment sizes that the illumina platform is capable of (I believe this is 100 bp to 600 bp)? If the answer is no then what are the disadvantages to doing this?
Any insight would be greatly appreciated.
Cheers,
Eager Scientist
This is my first post in this forum. Hooray!! Anyways, on to my question:
My lab is just beginning to get into RNA sequencing and we are currently working on our library preparation technique for sequencing using the illumina platform (GA2). In my travels through the literature on this topic I have noticed that several papers have suggested that during the post-amplification size selection of the library preparation, the band size should be relatively narrow. In other words if you are sequencing with 300 bp fragments it is better to choose a range of 300bp +/-50bp as opposed to +/-100 bp. Why is this? Is it okay to size select for the full range of fragment sizes that the illumina platform is capable of (I believe this is 100 bp to 600 bp)? If the answer is no then what are the disadvantages to doing this?
Any insight would be greatly appreciated.
Cheers,
Eager Scientist
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