Dear all,
I performed DNA extraction using Qiagen DNeasy Blood and Tissue Kit, followed the protocol with an addition of 2ul RNase A with incubation of 2 min at R.T after adding proteinase K. After that, I followed the protocol again.
I am going to send my samples for 16S Amplicon Sequencing and I am wondering if these are okay to send. And whether I need to treat them with RNase again before sending for sequencing.
For quantification, I need at least 400ng or 20ng/uL and strongly recommended more than 800ng with OD260/280 1.8-2.0.
For my samples, the quantification was done using nanodrop and are below;
1: 279.2 ng/uL, OD: 2.04
2: 146.9 ng/uL, OD 1.99
3: 249.9 ng/uL, OD :2.00
4:237.7 ng/ uL, OD: 2.03
5: 193.0 ng/uL, OD: 2.02
6: 392.4 ng/uL, OD 2.02
7: 352.5 ng/uL, OD: 1.99
Hello Andrew Ting
i Recommend only sample number on the well hel number 4, 5, 6, and 7 you can proceed for sequencing. Other samples are showing low visible band indicates you doesn‘t has enough amplicon for sequencing that samples (number 1, 2, and 3)
Andrew Ting
The Samples No 4,5,6 and 7 are good to go. 1,2 are too low in band visibility. send only 4,5,6,7 for sequencing, I doubt if 1,2 and 3 can come out with any results.
I recommend to proceed samples 4, 5, 6, and 7 for sequencing, while samples (number 1, 2, and 3) not good as it is, but you can duplicate it to be good enough for sequencing Andrew Ting
This is your genomic DNA prep. 16S amplicon sequencing will use this as template for PCR. All of your samples will amplify the 16S. So, go ahead, dilute them as per requirement and send for sequencing.
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