We have had happy experience with 454, pgm and illumina libraries. However, when we followed the minION Lambda DNA control library protocol, we can only get 25 ng DNA instead of the expected 250 ng.
First of all, the provided lambda DNA cannot be sheared to ~ 6kb to 8k even at 16,000 X g using g-tube. This is unusual based on our experience and Covaris protocol. The final sizes of the sheared DNA are around 12-15 kb. The support team told us no need to worry about the size...... After end-prep process, the yield of DNA is 300 ng instead of the expected 700 ng (30% instead of 70%). The final yield of the library is even worse, around 25 ng (8.3% instead of 35% ).
Two posts in the Nanopore community showed that the provided lambda DNA cannot be sheared to 8 kb. They got 16Kb. But nobody replied to those two posts...
Can we load the pooled ~ 60 ng library to minION? Any suggestions will be helpful. Thank you very much in advance!
First of all, the provided lambda DNA cannot be sheared to ~ 6kb to 8k even at 16,000 X g using g-tube. This is unusual based on our experience and Covaris protocol. The final sizes of the sheared DNA are around 12-15 kb. The support team told us no need to worry about the size...... After end-prep process, the yield of DNA is 300 ng instead of the expected 700 ng (30% instead of 70%). The final yield of the library is even worse, around 25 ng (8.3% instead of 35% ).
Two posts in the Nanopore community showed that the provided lambda DNA cannot be sheared to 8 kb. They got 16Kb. But nobody replied to those two posts...
Can we load the pooled ~ 60 ng library to minION? Any suggestions will be helpful. Thank you very much in advance!
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