Can anyone help me how to get the similar bands in Actin?

I run western blot on HCT-116 cell lines, I got actin band like in the attached picture. The band size is not same. Can anyone help me explain this situation and how to overcome? Thank you your...

22 February 2021 6,937 3 View

Can someone help me to improve the MCF-10A growth?

18 January 2021 7,640 3 View

How big the Balb/c mice spleen after immunization?

I am working with Balb/c mice, and in my experiment, I have to inject mice with antigens, then get their spleen and collect the splenocytes for fusion. Recently, I get into trouble with the mouse...

04 January 2021 3,591 3 View

Capture of oleophilic solid polymers present in water solution ?

I am looking for articles, coferences, publications or thesis concerning the capture of solid oleophilic polymers in solution in water. Thanks Thierry Le Blan

01 January 2021 5,798 3 View

Hybridoma stops producing antibodies after thawing from liquid nitrogen?

Dear scholars, Our research is currently involved with hybridoma technology. We have isolated some original clones after fusion and we are preparing the subclone step to find some true positive...

01 December 2020 1,178 3 View

Suggestion for normal ovary cell lines?

Dear all, I want to choose a normal ovary cell line and check for its proliferation property when introducing carcinogenic factors. Does anyone have any suggestions for the cell line? Many thanks...

08 November 2020 9,100 3 View

What is Snapback Primer in realtime PCR?

Dear all, I find a paper using Snapback primer. I do not understand the function of snapback tail? What is snapback primer? When do we use snapback primer? and why we use that? Thank you.

01 November 2020 8,869 2 View

Rotation matrix - 3D dimension to 2D dimension?

22 October 2020 9,354 4 View

Am I introducing any potential loss of RNA integrity by combining an RNALater solution with a non-ionic detergent?

I am planning to characterise the transcriptome of pre-anthesis kiwifruit anthers. The samples are being harvested on my behalf and sent down by post, and I cannot ask the harvesters to dissect...

10 October 2020 1,830 3 View

Why we got instability results when simulating elastic waves (Lamb waves) at high frequency using Abaqus?

Hi, I am trying to model Lamb waves propagation in an Aluminum plate using Abaqus. If I choose the frequency of 5-cycle Hanning excitation source is 0.2 MHz (to make sure that only 2 modes S0 and...

18 August 2020 7,022 8 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

Can someone suggest a way to prepare slides to observe under confocal microscope for observing fluorescent tagged proteins??

I am growing the cells on coverslips and transfecting the fluorescent tagged protein directly on them. Then i want to observe these cells under confocal microscope. Currently i am just using...

01 March 2021 6,142 3 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

Does RNAiMAX transfection reagent work in serum free medium?

I diluted siRNA and RNAiMAX in opti-MEM and added to the cells which they were in the growth medium. Is it a right way? or should I culture cells in the opti-MEM medium for a while and not in...

26 February 2021 10,041 3 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