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 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

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

Help me conduct a neuroscience experiment for my master's thesis?

22 January 2021 9,994 5 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

Anybody to help me with Code source for DOUBLE CONCAVE FRICTION PENDULUM BEARINGS Please?

Hello, would you please share the codes for Base Isolation of structures using Double Concave Friction Pendulum Bearings? I am doing a related research but as I am just a beginner in MATLAB, I...

18 January 2021 6,683 1 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

How can I verify my centrifugal fan design for electric motor cooling?

I have radial blade centrifugal fan design which will be mounted on exhaust side of electric traction motor. The fan will be mounted on same motor shaft inside the housing. How can I verify that...

23 February 2021 2,390 3 View

Are ZnS nanoparticles moisture sensitive?

Hello Everyone, I tried to form ZnS nanoparticles using the hot injection method and was able to form the same (confirmed by XRD). When I centrifuged, after the reaction I got a kind of white...

17 February 2021 1,709 3 View

Peptide precipitation as pre-step for acid hydrolysis for amino acid analysis?

I'm currently experiencing problems during acid hydrolysis for total amino acid analysis as my first samples contain high amounts of sugar (I get too low results back). I therefore had the idea to...

14 February 2021 6,049 3 View

What is the optimal propagation regime for pET-1M(KanR) and pMCSG7(AmpR) plasmids in DH5alpha E.coli?

Dear all, I am trying to harvest the low-expression plasmids in the mentioned E.coli strain. I constantly gel very low concentrations (30-50 ng/uL while the concentration is usually around 100...

08 February 2021 4,255 4 View

Why do we wash azolectin with acetone?

I want to prepare liposomes made of azolectin (sometimes spelled as asolectin). I start with the azolectin powder, and many protocols say that I should wash it with acetone (dissolve, centrifuge,...

04 February 2021 2,738 3 View

Difficulties purifying Outer Membrane Vesicles using a Sucrose Density Gradient: Unable to detect any protein after this process?

I'm currently trying to purify Outer Membrane Vesicles (OMVs) following their extraction from the bacteria I'm working with. After the initial extraction the concentration of protein seems good...

27 January 2021 6,852 3 View

How to get the insoluble fraction of PK digestion for mitochondria?

I am trying to check for some proteins if they are located in the inner or outer mitochondria membrane after treating the cells with a neurotoxin. I am using PK to digest the mitochondria, but I...

27 January 2021 5,145 2 View

What time and speed should the centrifuge be set at in order for cell debris and clumped proteins to form a pellet?

- The Eppendorf tube was then placed in a microcentrifuge. Notice how it is balanced with another tube placed opposite it in the centrifuge drum to ensure smooth rotation. Other information...

24 January 2021 2,547 3 View

SDS-PAGE aggregation problem?

I ran SDS-PAGE with 14% lower gel, 4 % upper gel, having brushing, pre-running empty gel for 30 min. The protein samples had been centrifuged with 14,000g for 10 min, and then the supernatants...

20 January 2021 172 3 View