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Similar Threads
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| Thread | Thread Starter | Forum | Replies | Last Post |
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12-14-2018, 04:22 AM
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#1 |
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Member
Location: Hanover, Germany Join Date: Sep 2018
Posts: 15
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ScriptSeq v2 135base-peak
Hello people,
I tried the ScriptSeq v2 RNA Library Prep Kit on the total RNA extract of a cerebrospinal fluid sample. The RNA amount was not measureable with our Qubit but I tried anyway. I followed the protocol, did the AMPure bead purification, ran 16 PCR cyles and got this massive 135b-peak in the bioanalyzer that towers everything. Any idea what this could be? I'm super new to RNA seq and not yet familiar with all the intricacies and pitfalls of the steps, so any help is appreciated  Cheers! |
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12-14-2018, 06:22 AM
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#2 |
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Senior Member
Location: Bethesda MD Join Date: Oct 2009
Posts: 509
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Looks like adapter dimer.
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12-15-2018, 02:39 AM
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#3 |
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Jafar Jabbari
Location: Melbourne Join Date: Jan 2013
Posts: 1,238
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Bead clean up with 0.8x should remove it. It may be necessary to do two clean ups for complete removal. Clean ups should be done with all samples that will be compared if the aim is DGE analysis.
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12-17-2018, 02:15 AM
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#4 | |
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Member
Location: Hanover, Germany Join Date: Sep 2018
Posts: 15
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Quote:
And thanks, I will try a 0.8x cleanup before sequencing. |
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12-17-2018, 11:31 PM
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#5 |
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Senior Member
Location: US Join Date: Dec 2010
Posts: 452
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Randomprimer-dimers will be basically unavoidable for your low, unmeasurable input.
If the oligos are a separate reagent, you could try to reduce their concentration. |
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| Tags |
| bioanalyzer, library prep, rnaseq, scriptseq |
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