Brandon:LabNotes/Project1/2015-7-24
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NXTA_ME_B HiSeq, MiSeq troubleshooting, libraries that worked on HiSeq on MiSeq[edit]
- size selection on samples from Tn5-059 versus ez-tn5 with THS-seq, using 500 cells and guanidine HCl method, since ran out of sample that was used for HisSeq.
- gel size selection amounts (see 7/24/15 size selection column on far right)
gel size selection amounts | ' | ' | ' | ' | ' |
4/2/2015 | intensity | ng/ul | |||
7213.490215 | 5 | for 7/24/2015 gel size selection | |||
5410.760161 | 2.5 | for 80 ng | |||
1. idx49, t7-top2, 5X tn5-059, 500 cells rep1 | 19053.90057 | 21.42011006 | 3.734808075 | 7.46961615 | |
2. idx50, t7-top2, 5X tn5-059, 500 cells rep2 | 16376.87049 | 17.70764341 | 4.517823076 | 9.035646151 | |
3. idx51, t7-top2, 5X ez-tn5, 500 cells rep1 | 18371.57055 | 20.47386464 | 3.907420578 | 7.814841156 | |
4. idx52, t7-top2, 5X ez-tn5, 500 cells rep2 | 16685.3305 | 18.13541129 | 4.411259205 | 8.82251841 | |
5. idx53, t7-top2, 5X tn5-059, 6 ng pure DNA | 38879.07116 | 48.91337028 | 1.635544628 | 0.8 uL | 3.271089256 |
6. idx54, t7-top2, 5X ez-tn5, 6 ng pure DNA | 34297.93102 | 42.56031131 | 1.879685499 | 0.8 uL | 3.759370998 |
7. idx55, t7-top2, 5X tn5-059, NTC | -2643.330079 | -8.669296012 | -9.227969594 | 1 uL | -18.45593919 |
8. idx56, t7-top2, 5X ez-tn5, NTC | -1150.120034 | -6.59853389 | -12.1239053 | 1 uL | -24.2478106 |
- gel size selection results
File:ZhangLab 2 2015-07-24 11hr 11min-labeled.jpg
- gel size selection validation
File:ZhangLab 2 2015-07-24 14hr 04min-labeled.jpg
Results[edit]
- samples did not work well on MiSeq, and did the same thing as other samples, with very low read numbers. Thus must be MiSeq problem.
- loading?
- special miseq reagents being used?