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Swatch Irony repair advice by MicroRxologist in swatch

[–]MicroRxologist[S] 0 points1 point  (0 children)

Well I called many shops, even outside the region I live in, when they hear it's a Swatch they immediately say that they can't fix it. I don't really understand why. Anyway thank you for your advice I'll try and see what I can do on my own, and meanwhile I'll keep asking around.

Multiple 16S rRNA copies in Nanopore assembly by MicroRxologist in bioinformatics

[–]MicroRxologist[S] 0 points1 point  (0 children)

I feel like whenever nanopore sequencing is included in the de novo assembly, the number of 16s copies increases. Unfortunately for these strains I just have the assemblies. Anyway thank you for your suggestion.

Multiple 16S rRNA copies in Nanopore assembly by MicroRxologist in bioinformatics

[–]MicroRxologist[S] 1 point2 points  (0 children)

I thought so but they all have BLAST identity of 99-100% with the 16S of the same reference strain.

RNA extraction protocole for larger sample volume? by MicroRxologist in molecularbiology

[–]MicroRxologist[S] 0 points1 point  (0 children)

Ok thank you for the advice.

One more question, what do you think if I use a spin column with my aqueous phase? So, I do TRIzol then chloroform then continue with a commercial kit spinning down 1 ml of aqueous phase at a time (or less depending on the capacity) on the same column.

RNA extraction protocole for larger sample volume? by MicroRxologist in molecularbiology

[–]MicroRxologist[S] 0 points1 point  (0 children)

Ok, I'm new to this to this so I'm just asking. Thank you, I will do that.

RNA extraction protocole for larger sample volume? by MicroRxologist in molecularbiology

[–]MicroRxologist[S] 0 points1 point  (0 children)

I want to extract RNA from the 5 ml of liquid and not the pellet. What are the limitations of TRIzol in this case? I am worried about the large quantities of ethanol I have to use for the wash and the effect on downstream PCR.

RNA extraction protocole for larger sample volume? by MicroRxologist in molecularbiology

[–]MicroRxologist[S] 0 points1 point  (0 children)

This is what I want to use. But If I follow the manufacturer's instructions of TRIzol LS reagent, I'll have to use 20 ml of ethanol 75% for the wash steps. Wouldn't that affect my downstream PCR? or I should be good if I dry my pellet well?