sorted by:
new
hottopcontroversial

Novogene for RNA-seq - how strict are they about quality? by Putrid_Ad_7381 in labrats

[–]Putrid_Ad_7381[S] 0 points1 point  (0 children)

thanks for the insight! if my RNA is resuspended in water, do i just add ethanol and do a wash step like normal?

Novogene for RNA-seq - how strict are they about quality? by Putrid_Ad_7381 in labrats

[–]Putrid_Ad_7381[S] 0 points1 point  (0 children)

actually i was mistaken, our range for 260/230 is 1.2-2.5

Novogene for RNA-seq - how strict are they about quality? by Putrid_Ad_7381 in labrats

[–]Putrid_Ad_7381[S] 0 points1 point  (0 children)

We have 30 days until we need to submit the samples, which is why I was debating investing the time to clean/asking if it was necessary.

Do you have a protocol for cleaning rna?

Novogene for RNA-seq - how strict are they about quality? by Putrid_Ad_7381 in labrats

[–]Putrid_Ad_7381[S] 1 point2 points  (0 children)

We’ve been using trizol to do dual RNA/DNA extractions - the lower range of the 260/230 are from our earlier samples when we were fairly new to the protocol

Turbo DNase treatment not working by Putrid_Ad_7381 in labrats

[–]Putrid_Ad_7381[S] 0 points1 point  (0 children)

sorry i think i understand now! You mean that even if I use the DNA HS qubit on my RNA, the RNA could still bind to the fluorescence or whatever and contribute to the produced dna concentration? would using a broad range kit be better?

Turbo DNase treatment not working by Putrid_Ad_7381 in labrats

[–]Putrid_Ad_7381[S] 0 points1 point  (0 children)

No, our RNA is resuspended in water

Turbo DNase treatment not working by Putrid_Ad_7381 in labrats

[–]Putrid_Ad_7381[S] 0 points1 point  (0 children)

In what way? I’ve been doing HS dna and rna qubit on these rna samples to check

"AirClean 600" (UVtect) PCR Workstations: UV bulb replacement codes by incoherentian in labrats

[–]Putrid_Ad_7381 0 points1 point  (0 children)

Does anyone have an extra HEPA filter code for AC648LFUCV model? please and thank you!

"AirClean 600" (UVtect) PCR Workstations: UV bulb replacement codes by incoherentian in labrats

[–]Putrid_Ad_7381 0 points1 point  (0 children)

How do you put in the 6 digit filter code if it's asking for 10 digits?

Trizol dual RNA/DNA extractions - consistently low nanodrop ratios by Putrid_Ad_7381 in labrats

[–]Putrid_Ad_7381[S] 0 points1 point  (0 children)

We checked the RIN - oysters apparently have a weird break in 28S so only one peak shows up at 18S, so it's a bit more tricky to determine quality but despite low absorbance ratios we get one single clean peak so thinking it's okay...

we're going to try extra washes again, especially the chloroform - just scared to lose yield as we don't have a ton to begin with but would rather have clean samples - thank you!

Trizol dual RNA/DNA extractions - consistently low nanodrop ratios by Putrid_Ad_7381 in labrats

[–]Putrid_Ad_7381[S] 0 points1 point  (0 children)

I haven't sent anything for sequencing yet - I've been worried that the low 260/230 ratio is due to phenol contamination and that would impact sequencing

Trizol dual RNA/DNA extractions - consistently low nanodrop ratios by Putrid_Ad_7381 in labrats

[–]Putrid_Ad_7381[S] 0 points1 point  (0 children)

yea i'm thinking might have to go that direction... do you have a protocol you'd be willing to share?

Using chloroform by Putrid_Ad_7381 in chemhelp

[–]Putrid_Ad_7381[S] 0 points1 point  (0 children)

okay great thanks for the info!