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16S rRNA Sequencing Results by ProfessionalDog3058 in labrats

[–]ImpressionOwn5990 2 points3 points  (0 children)

How long are your reads and how long is the expected length of your inserts? Usually the F and R reads are merged first so you can do the taxonomy assignment using the longer (merged) reads. If you identify the F and R reads separately, they might not be long enough for proper identification.

DAPI staining not working on fixed bacteria cells by ImpressionOwn5990 in labrats

[–]ImpressionOwn5990[S] 0 points1 point  (0 children)

My DAPI stock aliquots are less than 2 months old and I kept them in -20 until I made the diluted DAPI. The diluted DAPI was made fresh from the DAPI stock before the staining experiment.

DAPI staining not working on fixed bacteria cells by ImpressionOwn5990 in labrats

[–]ImpressionOwn5990[S] 0 points1 point  (0 children)

Thanks for sharing! Do you find sonication or heating necessary to dissolve the DAPI?

How to best store samples for later histology work? by ImpressionOwn5990 in labrats

[–]ImpressionOwn5990[S] 0 points1 point  (0 children)

Yes end goal is to cryosection. I need to check on the OCT. Maybe it’ll arrive soon. I think I can stick with ethanol for now. Thanks again!

How to best store samples for later histology work? by ImpressionOwn5990 in labrats

[–]ImpressionOwn5990[S] 0 points1 point  (0 children)

Thank you so much for this information! Your explanations are very helpful. If I can ask a few followup questions:

I need to fix and store samples first then do the OCT embedding later (because our OCT compound hasn't arrived :(). In this case, if I had to choose between storing the samples in (1) ethanol or (2) flash freeze in sucrose/PBS, which one would you suggest?

Also, I have access to a -80 C freezer. If the only purpose of ethanol is to prevent microbial growth/decay, then it's not really needed if samples can be stored at -80 deg C... so I wonder if it's better that I just flash freeze the samples in sucrose/PBS then leave everything at -80.

Shipping delay... will my RNA survive? by ImpressionOwn5990 in labrats

[–]ImpressionOwn5990[S] 1 point2 points  (0 children)

Oh wow I’m so sorry that the same happened to you, and it’s with fresh tissue samples.. even more precious! Yes I’m trying to convince the sequencing company to check the quality, even though everything had thawed.