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

Will the plasmid without SV40 Ori have a quite low expression level in 293T?

SV40 Ori helps plasmid to replicate in 293T cells. If I use a plasmid which doesn't have a SV40 Ori, will the expression level be very low?

10 December 2020 1,751 2 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

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

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

How vaccines of covid 19 work?

Millions ask how vaccines of covid 19 be successful and safe, its here to explain generally how vaccine of pfizer work ,mRNA vaccines teach our cells how to make a protein that will trigger an...

28 February 2021 2,723 4 View

What is the most suitable server for predicting protein structures in bacteria?

I have used the i-Tasser several times, however it has been unavailable for several days. I tried the swiss-model, but the output was not very pleasant due to the model used. Is there any other...

28 February 2021 4,521 3 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

Are protein samples in SDS compatible with ELISA?

I have used an AllPrep DNA/RNA/Protein Mini Kit QIAGEN kit to extract RNA and protein from my samples. At the end of the protein purification, I resuspend my protein in 5% SDS. Will these samples...

28 February 2021 7,370 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

How to replicate the well conditions of a crystal hit with 20%(w/v) isopropanol?

I had a crystal hit in a well with 0.1M HEPEs pH 7.5, 10%(w/v) PEG 4000, and 20%(w/v) isopropanol. Why is isopropanol in w/v here if it's just solvent? How do I recreate this well condition?

25 February 2021 9,761 1 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

What urea buffer can i use for biotagged protein extraction from streptavidin beads?

Hi all, I have been doing research on biotagged LDB1 proteins in Mel cells. I purify the proteins using M280 streptavidin beads. The yield is very low so I've been trying to optimize it by...

24 February 2021 5,749 3 View