Does entering wrong information in rt pcr machine affects the results?

I wonder if entering a wrong information to rtpcr machine would really affect the results. For instance, wrong concentration or dilution factor when labeling the plate before the run.???

23 December 2015 725 1 View

Why do I get double bands on gel electrophoresis for rtpcr standards?

17 November 2015 4,188 2 View

Does rtpcr results influnced by the cdna concentration and 260/280 ratio?

I have cDNA samples with concentrations range from 99- 6700 ng/ul and 260/280 ratio of 1.5- 2.02. I run a rtpcr using some samples to conduct the standard curve, however, the -ve (blank) curve was...

14 October 2015 7,866 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

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

The reason that the culture of white fungi Contamination with green mold(Trichoderma), the culture autoclave at121 for 15 min. What is it?

Is the period to autoclave not enough? The inoculation in a hood and flame and UV and alcohol.

27 February 2021 9,356 3 View

No amplification data for qPCR on MxPro?

I have set up a qPCR run on MxPro with SYBR green and ROX as a reference, however, when I go to the analysis section the program seems to only collect data for the dissociation curves and not the...

25 February 2021 8,039 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

Color of spiro-ometad film?

What does the color of a spiro-ometad film indicate? I got purple, green, red, even black color of the spiro film. Is there anything we can get from the preliminary color analysis(e.g. oxidation...

21 February 2021 1,831 4 View

Has anyone encountered any problems with abm good microRNA primers for quantification of mature miRNAs?

I've been trying to quantitate mature microRNAs in 293T cells with and without 'X'. 2ug of total RNA (isolated by Trizol method) is converted to cDNA using All-in one microRNA cDNA synthesis kit...

17 February 2021 8,986 2 View

Does the property of GFP affect a fusion protein's localization?

Hello, I am planning to insert GOI between CaMV 35 promoter and mGFP in pCambia1302 to produce a fusion protein and see subcellular localization of the protein. But the vector says it's membrane...

15 February 2021 4,353 3 View

How do you prepare a standard dilution for qPCR?

I am very new to quantitative PCR and need some help for this technique. For my experiment, I need to perform a quantitative reverse transcription PCR. I extract the RNA from my pellet and...

05 February 2021 9,834 4 View

P53 detection in methylated and non methylated samples?

Hello everyone. I am about to search for the expression of genes including p53 in methylated and non methylated HPV samples. Should I extract DNA from the methylated samples and use SYBR green...

04 February 2021 2,749 3 View