QPCR troubleshooting - what could cause an increasing ROX signal?

Hi everyone, I'm looking for some help troubleshooting a recent qPCR issue I've encountered. I am using fluorescent Taqman probes and a qPCR master mix that includes ROX as a passive reference....

08 February 2021 419 3 View

How to isolate RNA in a way that is representative of an entire brain (fish)?

Hi there, I'm looking to compare the expression of a particular gene in fish brains across three developmental stages. My aim (as first step) is to use the entire brain for RNA isolation steps....

02 September 2020 7,195 3 View

Author unable to be contacted prior to publication?

Can anyone let me know what to do in the case of a key investigator being uncontactable at the time of manuscript submission. Most journals ask for a signature and address of all authors and yet...

25 January 2018 1,627 3 View

Can I load samples into an electrophoresis gel, let sit overnight, & then turn the power supply on in the morning (and the gel run smoothly)?

samples (from yeast)= Whole cells, spheroplasts, crude mitochondria, and isolated mitochondria

24 August 2017 3,677 16 View

Trouble with Phosphorylation/Ligation of a blunt-ended CRISPR plasmid. Any troubleshooting ideas?

I have been trying to phosphorylate and ligate a blunt-ended linear plasmid after PCR. It remains linear. Any suggestions for troubleshooting? I have tried 10 min at 70 degrees followed by ice for...

15 March 2017 3,701 3 View

How do I do hierarchical cluster analysis in STATA on 11 binary variables?

I have 11 sleep outcomes - all binary .i.e. initial insomnia? yes/no. I want to see whether the 126 individuals I have cluster onto the different sleep variables. I have tried EFA before, and the...

28 February 2017 9,569 3 View

I'm trying to silence a microRNA using an antagomiR and lipofectamine2000 and have not been successful so far. Does anyone have any tips?

19 June 2016 5,496 6 View

In C57/Bl6 immunisations/challenges, are there any differences in using CD95 or GL7 positivity as a marker for germinal centre B cells?

06 October 2015 2,298 2 View

How long can I incubate C3H10T1/2 cells (or 3T3-L1 cells) with cell permeable PKA inhibitor 14-22 amide?

And should I incubate them at 37C or room temperature? If at RT, should it be in the dark? I am differentiating the cells into adipocytes, using the MDI protocol (IBMX, dexamethasone and insulin)...

30 September 2014 8,930 2 View

Interested to stain mice tissue with WGA 488 conjugate, anybody has a protocol to recommend?

Interested to stain mice brain tissue with WGA 488 conjugate, appreciate if anyone willing to offer me some guidance or suggestions! Thank you!

28 February 2021 3,951 2 View

Can anyone suggest a way to dissolve insulin which is for rat/mouse injection?

We are trying to inject insulin to rat or mouse, but the insulin we've got is powder, and there are many ways for dissolve, like in saline, PBS, DMSO, or in Tris-HCl, can anyone give us a...

13 February 2021 8,426 10 View

Cell culture compatible glue/sealant?

Hello We are looking for a glue /sealant to fix and secure some glass parts of a live cell imaging chamber. It has to be biologically inert because it may come in contact with the medium, and...

01 February 2021 6,886 3 View

Does anyone have a copy of the sTOFHLA?

Peppercorn books is out of business, and that the creator of the test has been dead for over 5 years. I am looking to use this tool to assess literacy in community dwelling seniors as part of my...

25 January 2021 4,061 2 View

Can Cas13a protein directly purify from PAGE gel, please suggest me the method?

I am willing to Cas13 in vitro. Can it be directly purified from PAGE gel, please suggest me the method.

19 January 2021 1,136 3 View

Is there no general protocol for vaccination study?

Hi, I searched vaccination study to know which MLD50 is suitable for influenza virus study in pubmed. But why every vaccination and challenge conditions are different? Is there no guideline or...

16 January 2021 7,984 3 View

Can data collected through participant observation be interpreted through Interpretative phenomenology?

I have just started to read about phenomenology and hence quite a beginner. I have observed hospital interactions between doctors and patients, noted down the conversations, added the non-verbal...

14 January 2021 3,452 3 View

Johnson et al., 2006 Geology of South Africa?

I am wondering if there is a digital (PDF) version of this book available or if anyone is willing to share this work. It is no longer available from GSSA (Geo Soc. S. Afr) nor does my library hold...

13 January 2021 5,571 2 View

Will there be two research designs if using Mixed Methods?

I am having a hard time with my dissertation pre-proposal (super long story, but I have no help). If I use the Mixed Method, will my research design will be both phenomenological and descriptive...

07 January 2021 5,007 3 View

Do you think that genetic inheritance or the environment is more influential in the development of substance abuse?

Do you think that genetic inheritance or the environment is more influential in the development of substance abuse ?

04 January 2021 1,587 3 View