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Originally posted by BTS View PostThere is some very useful information here. Has anyone used Bio-rad's Gel Doc EZ system in place of a bioanalyzer?
The Gel Doc EZ system is good for taking pics of gels and doing other work but you can't beat the bioanalyzer's ability to handle different sample types like the High Sensitivity/DNA1000 chips for DNA and nano/pico for RNA. We use it multiple times a day for sample QC -> QC through library preps.
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Originally posted by NextGenSeq View PostIf you use Q-PCR to quantify your libraries you don't need a Qubit. A NanoDrop is still useful though, as well as a Bioanalyzer.
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There is some very useful information here. Has anyone used Bio-rad's Gel Doc EZ system in place of a bioanalyzer?
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Originally posted by GW_OK View PostHere is my list, in decreasing order of indispensability:
Bioanalyzer
qPCR instrument
Covaris
Qubit
Robot (We've got a SPRI-TE)
Size selection system (We don't have one, but if you're robot can't size select for you it might be nice)
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Here is my list, in decreasing order of indispensability:
Bioanalyzer
qPCR instrument
Covaris
Qubit
Robot (We've got a SPRI-TE)
Size selection system (We don't have one, but if you're robot can't size select for you it might be nice)
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I would agre with the Bioanalyser and Qubit. Standard manual gels are fine, if run carefuly so as to reduce variability between gels.
The Bioruptor is an alternative to the Covaris.
I would also recommend a disposable, gel cutting tips. These are indespensible in gettting the same amount of gel/DNA into the ceanup column.
You might also consider something like Beckmans SPIworks or a Tecan, Beckman or even Illumina robot to automate the TruSeq library kits.
Pippin Prep and Calipers system are both eary days instruments and it wil be interesting to see how they work out compared to manual or SPI based selections.
Have fun.
Oh, and dont forget the largest stress relieveing device you can find!
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Thanks for the input! Does anyone know if the Qubit uses RiboGreen for quantification, or is that another Fluorometer? From everyone's experience, which do you prefer based on time/reliability/sensitivity/cost: Qubit/qPCR/bioanalyzer?
I also haven't done much with e-gel, but hear that the pippin prep is quite nice for separation & size selection. Has anyone used both? Could you get away with standard agarose gels if needed and the pippin prep for 'gel extractions' & size selection?
What model Covaris do most people use? S2?
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If you use Q-PCR to quantify your libraries you don't need a Qubit. A NanoDrop is still useful though, as well as a Bioanalyzer.
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I agree with the previous answers, and would probably add the Covaris as well.
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We also have a Qubit. Very useful and quite inexpensive to buy and use. A nanodrop is also required for SOLiD long mate pair. Try and get use of someone elses as we dont use a nanodrop for anything else but circularised DNA.
If you are not going to use SPRI beads get an e-gel setup. 10 to 15 minutes rather than 30mins to 1 1/2 hours for a normal gel.
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