PACYC PCR not working after several attempts. Would anyone have any suggestions?

So, I have been trying to run a pACYC PCR which will be used later on for a Gibson Assembly. However the PCR is not working. I have already tried gradient PCR and changing extension time; however...

02 March 2021 1,146 2 View

Quantifying leaking DNA from bacteria - what's the secret?

Hi, all! I need to determine if my treatment causes membrane damage in bacteria (S. aureus and E. cloacae), but I cannot use a fluorescent probe such as PI and similar ones because my compound is...

24 February 2021 2,006 2 View

Please let me know how to remove mycoplasma contamination from cell line?

I am facing a problem in maintaining the cell line of HepG2 and HepG2.2.15. I have used plasmocin Tm treatment (Invivo gen) Cat.#ant-mpt Lot #MPT-41-01 for more than a month. I have also used...

21 February 2021 6,046 4 View

Western blot protein of interest falling at a lower weight than expected?

My protein of interest is DAGLa (115 kDa). We received the antibody for this from another researcher who reliably shows bands at this weight. We do see a light banding at this weight, but the...

11 February 2021 6,009 3 View

Which anesthetic for cortisol determination in fish?

I am a bit confused. I am planning an experiment in the field where fish will be caught by electrofishing. Afterwards, they are supposed to be euthanised by using an anaesthetic. I found...

10 February 2021 1,101 4 View

Which Journals would you advise to submit a Letter which briefly talks about neurosurgical treatment of neuro-oncological patient during Sars-COV-2?

Possibily not an open access journal which does not add any publishing costs once being accepted. Thank you in advance

09 February 2021 717 6 View

Can I modify vermiculite or perlite by adding nutrients to control a certain situation?

I need to assimilate a determining condition for the plant. However, I need to ask you if it is possible to change the properties and concentration of plant nutrients in this material like perlite...

04 February 2021 8,616 11 View

Sample Size for Quasi-Experimental Quants using one-tailed t-test?

Hi, sorry I'm stumped. I'm going to do a study on the leadership styles (using the MLQ Form 5X) of minority women administrators in medical schools. How will I calculate the sample size? Thank...

30 January 2021 2,865 4 View

Can you do Ponceau staining after loading control for Western Blot?

Hello everyone! I forgot to take a picture of my Ponceau stain prior to starting my first round of primary and secondary antibodies. I'm currently doing my loading control now and was wondering...

25 January 2021 757 3 View

What is the correct way to apply a Design effect for sample size calculation when the sampling method is linear systematic random sampling?

When we are doing sampling by means of simple systematic random sampling,how do we calculate the design effect for use during calculation of sample size for our work?

20 January 2021 5,328 3 View

Buffer to disrupt platelet alpha granules for PF4 quantification?

Hello! I want to quantify by ELISA the secreted (from platelets poor plasma) and the non-secreted (from platelet lysate) PF4 before and after TRAP stimulation. I will use the ELISA from R&D...

03 March 2021 1,499 2 View

How to grow E.coli on the low salt LB agar plate with 50 or 100ug/ml of Blasticidin?

Hello, I would like to ask how to grow E.coli which resists 50 or 100ug/ml of Blasticidin. I have tried many methods by using low salt Luria-Bertain(LB) agar plate. However, no colonies grow. Let...

01 March 2021 2,027 2 View

What are the countries most affected by extreme rainfall as a result of climate change?

Increased heat, drought and insect outbreaks, all associated to climate change, have increased wildfires. Decreasing water supplies, declined agricultural yields, health impacts in cities owing to...

01 March 2021 7,168 9 View

Dapi fluorescence protocol for colon cell lines ?

28 February 2021 2,534 2 View

Can bacterial suspension be pipetted directly onto petri dish and agar poured onto it?

Hi, recently I encountered a protocol for plating which was not the usual method I have been practicing (pour LB agar onto plate, allow it to solidify, pipette bacterial suspension onto plate,...

28 February 2021 7,292 3 View

Difficulty in cloning of 1kb gene into vector?

I am facing difficulties in cloning a 1kb gene into a vector (pJIT163). I have my gene on interest (GOI) in pUC57 and want to clone in pJIT163 using SalI and BamHI restriction sites. I am getting...

25 February 2021 3,221 7 View

Can I use acetic acid with an ELITE WAX Perkin Elmer column for Gas Chromatography?

Hi! I would want to know if I can use acetic acid with an ELITE WAX Perkin Elmer column for Gas Chromatography. This column has a polyethylene glycol phase, but I can't find the manual to check...

22 February 2021 730 2 View

How to seperate PEG-4000 from aqueuse phase ?

Hello everyone , Please tell me if It possible to seperat POlyethelyn glycol -4000 from an aqueuse media by other ways rather heating . I have tried salting out by electrolyte solution to...

22 February 2021 7,285 5 View

Can 5-FOA Diffuse Through Agar?

I am preparing to do a gene knockout in the filamentous fungi Coccidioides posadasii targeting pyrG/URA3 so that 5-FOA can be used for selection pressure instead of traditional antibiotic...

16 February 2021 169 4 View

Trouble shooting with his tagged protein expression in E.Coli?

i have issues in expressing a chicken protein in BL21 cells with his tag. I do not see induction even at 18 degrees. Any suggestions

15 February 2021 8,939 3 View