Kapa lib quant question

wei210 · 10-08-2019, 09:26 PM

Hello. I am new here. I have a naive question. When we make series dilution for kapa lib quant.
1) Will it make big difference if dilute the lib using H2O vs 10mM tris PH8.0 w 0.05% tween-20 ?
2) will kapa conc typically higher than the Qubit conc (lib w Illumina p5/p7 sequence primer) ?
3) vetex to mix should be better than Pipet up/down. In the case using Pipet up/down. How many times up/down should be sufficient?
Thank you all so much

seq198 · 10-10-2019, 02:11 AM

For PCR-free libs kapa quant kit makes sense, as qubit will not deliver accurate results. he different buffers for diluting should not have a big effect for what I can say. 5 times pipetting up and down should be sufficient.

wei210 · 10-10-2019, 06:18 AM

Thank you so much, Seq198

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10-08-2019, 09:26 PM	#1
wei210 Junior Member Location: Texas Join Date: Jul 2019 Posts: 4	Kapa lib quant question Hello. I am new here. I have a naive question. When we make series dilution for kapa lib quant. 1) Will it make big difference if dilute the lib using H2O vs 10mM tris PH8.0 w 0.05% tween-20 ? 2) will kapa conc typically higher than the Qubit conc (lib w Illumina p5/p7 sequence primer) ? 3) vetex to mix should be better than Pipet up/down. In the case using Pipet up/down. How many times up/down should be sufficient? Thank you all so much

10-10-2019, 02:11 AM	#2
seq198 Member Location: Germany Join Date: Dec 2015 Posts: 10	For PCR-free libs kapa quant kit makes sense, as qubit will not deliver accurate results. he different buffers for diluting should not have a big effect for what I can say. 5 times pipetting up and down should be sufficient.

10-10-2019, 06:18 AM	#3
wei210 Junior Member Location: Texas Join Date: Jul 2019 Posts: 4	Thank you so much, Seq198