Sam:LabNotes/Microbiome-new/2010-11-15
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2nd size selection of pooled Illumina library(short and long amplified E.coli SAGs -3rd test 11-07-'10)
Background
- Eight indexed indivudual libraries were pooled based on equal amount (20ng)
- The pool library will be size-selected again in order to get a narrower band for sequencing.
Procedures
- The pooled library is ~40uL. The pooled library was seperated into two tube (20uL/tube) for size-selection using 5-well TBE-PAGE.
- Sample well: 10uL H2O + 10uL 6X loading dye + 20uL DNA sample
- Ladder well: 30uL H2O + 10uL 6X loading dye + 0.5uL Low mass ladder/1kb plus ladder
- Run at 200V for 30min. Long staining (15min with foil wrap protection).
File:Sam111510-TBE PAGE.jpg File:Sam111510-TBE PAGE-post cut.jpg