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

Can I use breaking series as a regressor in a VAR model?

Context: I have two series, price and sales. Sales is mean-reverting stationery but price is stationary only after controlling for an intercept break. I want to set up a 2-equation VAR model...

14 January 2021 6,824 12 View

Does the use of Tissue Level implants reduce the risk of Peri-implantitis, as opposed to using bone level?

12 December 2020 6,007 6 View

How to calculate the normalized frequency V of photonic crystal fiber using COMSOL?In particular, how to calculate NEFF and nFSM using COMSOL?

24 November 2020 967 2 View

HOW to do an intention-to-treat analysis(ITTA)?

I mean, how do I put the dropped out participants' outcome (Missing value) in my analysis? Although I have already known what's ITTA meaning, I still don't know how to do it. Which methods should...

23 November 2020 9,982 5 View

Does pEGFP plasmid have G418 resistant?

Instruction from pEGFP: The recombinant EGFP vector can be transfected into mammalian cells using any standard transfection method. If required, stable transformants can be selected usingG418 ....

20 October 2020 4,662 2 View

The solubility of ethinyl estradiol (EE2) in methanol?

What is the solubility of ethinyl estradiol (EE2) in methanol? In the experiment, methanol was used to help dissolve, what is the reason for the flocs appearing after the storage solution is added...

20 September 2020 4,055 1 View

How to compare methylation status between different cell lines?

As in the title. I got the methylation status of several genes in different cell lines by bisulfite sequencing. More specifically, I have the sequencing data from the amplicons of these genes...

30 August 2020 6,020 2 View

How can I digest the formaldehyde (1%) crosslinked DNA-protein with restriction enzyme (DpnII) better?

I am working on yeast samples for 4C-seq study , but the formaldehyde fixed samples can not be digested with restriction enzyme (DpnII). Please help me out. Thanks.

28 August 2020 6,483 3 View

Where may I find a common ultrasonic image database for breast lesion's detection and segmentation, please give some suggestions?

to detect and segment breast lesion for breast cancer in ultrasonic breast imgae

27 July 2020 8,338 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

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

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

Problem with ligating 2.8 kb insert into 8.5 kb plasmid?

I am trying to clone 2 copies of the same mammalian gene into the pSF-CMV-Ub-Puro Ascl (contains HindIII, KpnI and NheI cut sites) plasmid. This is what the final product is supposed to look like...

24 February 2021 6,310 3 View

Best plasmid could be used as a standard curve for AAV titration?

I have been trying to have a good AAV titration for my production and I have tried different plasmids to set up as a standard curve but each one behaves differently. i am trying to use ITR for...

23 February 2021 5,167 3 View