Lentivirus particle production - Where's the integrase?

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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

What program can I use to caclulate dipole moment efficiently and without the need of excessive computation?

Hi, I am a third year chemistry undergrad doing my group project in which I need to predict the HPLC retention time of 20 aromatic molecules. to do this we require the chemical properties of these...

24 February 2021 1,384 3 View

Do you kow of any database containing data on retention time, log P and polarizability for a large number of molecules?

Hi, I am a chemistry undergraduate doing my dissertation project on HPLC. In my project, I need to predict the HPLC retention time of 20 aromatic molecules, the calibration set. To do this I will...

24 February 2021 2,602 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

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

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

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

What are the most relevant MSc proposal topics for Petroleum Production Engineering at the moment?

I am a Petroleum Production Engg MSc student at Robert Gordon University in Aberdeen and I am in the process of selecting a topic for my MSc Engineering Investigation proposal. I would very much...

01 March 2021 7,396 4 View

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 3 View

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View

PUC19 as a transfer plasmid for lentiviral delivery?

Hello, Is it possible to use pUC19 as a transfer vector to be packed in using the second generation viral particles packaging system( pMD2.G; psPAX2 plasmids)? As far as I understand it there is...

28 February 2021 4,868 2 View

Did anyone ever try using pLVX-TetOne Plasmid with a copGFP as a selection marker instead of Puromycin?

When using a lentiviral vector for inducible expression of genes using the Tet-On system. In the literature, I saw a widely used of pLVX-TetOne-Puro Plasmid, is there a reason why people prefer to...

25 February 2021 1,011 3 View

Has anyone had luck using cell lines other than HEK 293 (such as COS1) to package lentivirus?

I am looking to package lentivirus in endothelial cells but I am opposed to using 293 cells due to personal ethics regarding the source. I have done some reading and saw that COS1, COS7, & CHO...

25 February 2021 3,030 1 View

Why do the PCR bands from the cDNA templates (extraction from the cell after transfection) show different size from the band of the plasmid template?

After transfection with the plasmid ( linearized ) and subcloning of the cell lines, RNA was extracted from the cell and then reverse transcripted to cDNA. When PCR reactions were run to verify...

25 February 2021 5,712 3 View

How do I transfect a large plasmid into PC12 cells with nerve growth factor (NGF) receptor?

I have a large 16 kb plasmid, which I need to transfect into PC12 cells. Lipofectamin 2000 didn't work and with GFP alone the transfection rate is very low. I also tried the neon invitrogen...

25 February 2021 6,635 5 View