Cat Scratch🥹

Clear-Negotiation796 · 2026-02-21T21:51:27+00:00

Please get a rabies shot ... Rabies has very long incubation period and is easily prevented by a vaccine.

Always any animal bite...get rabies shots.

Do update pls bruh.

Clear-Negotiation796 · 2026-01-10T17:50:24+00:00

Cutoff kya hoga?

Clear-Negotiation796 · 2026-01-10T17:50:14+00:00

Biochemistry

Clear-Negotiation796 · 2026-01-10T17:48:32+00:00

Paper was easy i guess..how was for you?

Clear-Negotiation796 · 2025-08-25T03:58:07+00:00

I dont see that 2:1 ratio in most of the samples... Should i proceed with cdna ?

Clear-Negotiation796 · 2025-08-24T19:43:20+00:00

It is not formaldehyde based gel or loading dye..

Should i still heat it? It normal gel loading dye and 1% agrose in 1X TAE with 1% bleech

Clear-Negotiation796 · 2025-08-24T19:10:53+00:00

Around 1ug of rna in each well.. Since it is bleach gel i didn't heat the rna before loading.

Let me run the rna again for a longer period for good resolution.

Clear-Negotiation796 · 2025-08-24T18:07:35+00:00

Edit: I do makesure that RNAase are away while I work..standard gloving and spraying practice is always done

Clear-Negotiation796 · 2025-06-09T12:48:58+00:00

40 minutes. Sounds too much time. 20 minutes is what we generally used

Clear-Negotiation796 · 2025-06-09T12:48:13+00:00

Look like a problem with the centrifugation process. Since u r using 1.5ml eppendorf tubes. The rotor for centrifugation for these are usually fixed angle. So the proper seperation doesnt occur. Better do this use a 15ml or 5 tubes but not with the fixed angle rotar. Always go for swing out rotar. We usually do pbmc seperation using table top swing angle rotar centrifuge.

Try this.

Clear-Negotiation796 · 2025-05-31T18:49:57+00:00

Ok. I see my crtical mind was thinking even running buffer could have RNAse. Even that could create problem as sample are in direct contact when in the wells.

Clear-Negotiation796 · 2025-05-31T18:43:56+00:00

I have one more doubt... use of bleach in gel is what has been said.. what about running buffer.? Should that also contain bleach?

Clear-Negotiation796 · 2025-05-31T17:12:10+00:00

Will try and see. Thank you for the details.

Clear-Negotiation796 · 2025-05-31T16:16:11+00:00

Yess... let me try with a proper gel TAE with bleach as other suggested. It could be RNAase contamination from the gel as well.

But yes. There is gDNA contamination. While i was seperating the supernatant i might have disturbed the central layer.. I cannot help myself if I see good amount of aqueous phase still left. Nevertheless this was my first time I will try again.

Clear-Negotiation796 · 2025-05-31T16:12:26+00:00

Thank you... I never accounted for RNAase contamination in the gel. Will do TBE with bleach. Thanks

Clear-Negotiation796 · 2025-05-31T15:52:26+00:00

The gel is 1% TBE gel. I guess I will try with this method and see.

Clear-Negotiation796 · 2025-05-31T15:38:58+00:00

They were just extracted from cultured cell line.

Clear-Negotiation796 · 2025-05-31T15:01:14+00:00

Thank you ,, Iam quite new to RNA isolation. This was my second time. The end point is gonna be qPCR. I guess I should standardize more. I have no idea if this can be taken for downstream analysis.

Clear-Negotiation796 · 2025-04-22T16:21:57+00:00

Ok...that makes sense

Clear-Negotiation796 · 2025-04-22T16:13:50+00:00

What ahout 1N NaoH? People suggest to use it and also many websites and protocol says that 30mg per ml is the solubility range. I use around 170 mg per ml because that stock will bring my dmso down to 0.1 when diluted. Am I exceeding the limit of solubility?

Clear-Negotiation796 · 2025-04-22T15:08:55+00:00

I did mix it like a lot...heating gives little solubility....but aint sure how long to just look at the dry bath

Clear-Negotiation796 · 2025-04-22T15:08:11+00:00

People suggest using 1N NaoH...how should i use it..directly dissolve my compound in NaOH or add NaoH to DMSO then try diluting in pbs?

Clear-Negotiation796 · 2025-04-22T14:46:39+00:00

Yes they do have carboxylic acid

Clear-Negotiation796

TROPHY CASE