Cloning

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Questions related to Cloning

Is my protocol effective for extraction of protein from cell fractions?

Fractionation 1. Scrape cells and collect by centrifugation (500 × g, 5 min, 4 °C) 2. Resuspend cell pellets in lysis buffer and incubate on ice for 10 minutes 3. Centrifuge at 500 x g for 5 min...

26 March 2024 6,313 3 View

DNA gel does not showing bands? Troubleshooting help?

Bands are appearing very, very faint! I presume low DNA in gel, but my DNA concentration is over 1ug/ul. This is after my overnight digestion. I used a MaxiPrep protocol to extract DNA from...

26 March 2024 3,568 3 View

Xcrysden is displaying incorrect crystal structure and ibrav. But vesta identifies correct symmetries. Will it effect the end calculation in QE?

Currently i am working on a system of i4/mmm ( material composition is AB2C2) class Which corresponds to ibrav =7. I have sourced my CIF file from crystallographic open database, have crossed...

25 March 2024 363 3 View

Impact of sodium citrate pH 5 on cDNA synthesis and RT-PCR analysis?

Hi all! I can't dive into details, but I'm having issues with neutralising sodium citrate pH 5 in a 2-step cDNA synthesis and final RT-PCR. I'm manipulating my reverse transcriptase (100uL...

25 March 2024 7,186 0 View

How does nuclear fusion work?

What materials are used? What makes it so expensive?

22 March 2024 3,869 2 View

Any protocol for loading dsRNA in an agarose gel?

I am expressing dsRNA using E.coli as vector. I would like to check that my bacteria culture is expressing dsRNA. I am planning to extract RNA from my bacteria culture and then load the total RNA...

19 March 2024 1,351 3 View

Correct method for Short term Denaturated Inclusion body storage?

Hi, This is E. Park. I'm trying to purify a protein which was produced in E. coli as inclusion bodies. I already solubilized the inclusion bodies using a 6M guanidine-HCl buffer. Here is my...

19 March 2024 3,665 1 View

I don,t know why my TSI slant test false?

I do a TSI slant test, with Vibrio parahaemolyticus. Result of the test is A/A, but the correct result is K/A. Please explain this for me.

17 March 2024 8,852 0 View

I don,t know why my TSI slant test false?

I do a TSI slant test, with Vibrio parahaemolyticus. Result of the test is A/A, but the correct result is K/A. Please explain this for me.

17 March 2024 2,316 0 View

How disciminate différents species with qPCR ?

I'd like to detect the presence of different species in a DNA extract using qPCR. Are there specific targets already listed for each species (animals, yeasts)? Thanks,

14 March 2024 7,555 4 View

How microorganisms are important for maintaining of healthy soil and biodiversity and role of microbial communities in soil health and agriculture?

11 March 2024 1,435 6 View

Failure to deliver mRNA to T cells via LNP, how to solve this problem?

When we used LNP to deliver GFP mRNA to Jurkat cells, the efficiency was as high as 90%. However, when we delivered GFP mRNA to T cells, we failed, with an efficiency of 0.5-0.6%. Could anyone...

07 March 2024 3,307 2 View

Why the concentration of my expression vector drop with time and how to stop it ?

Hello everyone, I use several pcDNA3.1 expression vectors to transfect cells. The vectors were prepared by midi-prep a year ago and diluted in TE buffer. Now that I run new experiments, I...

05 March 2024 1,840 3 View

Why is my western blot not transfering straight?

I have human brain tissue samples that I lyse with a nuclear extraction buffer and then the pellet I solubilized in 8M urea. I run samples in Tris-Glycine buffer in premade 4-12% gels. Ran the gel...

05 March 2024 9,691 0 View

What is the optimal media and culture conditions for pituitary cells in vitro culture?

Dear Colleagues..I need any relevant publications/ technical procedure on the optimal method for pituitary cell, gonadotropes, in vitro culture....

29 February 2024 9,685 0 View

Does same gene dose mean same amount of expression?

My question is on the way of expression of allele specific copy number variation, e.g. individual 1 has a CNV as 3, 3, on each allele, with 6 copies; and individual 2 has 1, 5, on either allele,...

29 February 2024 1,563 1 View

I'm using the Purelink RNA Minikit for RNA extraction from whole blood. Can anyone advise on improving both yield and quality without switching kits?

I've been conducting RNA extraction from 200 microliters of whole blood using the Purelink RNA Minikit according to the provided instructions. Despite several attempts, I'm consistently obtaining...

23 February 2024 6,267 4 View

What is the price (average) in USD of Microwave-assisted hydrodistillator with a 1000 mL flask?

I need to buy this equipment to obtain several essential oils.

19 February 2024 353 0 View

Do you have an easy and clear reference enabling the good identification of arbuscular mycorrhizal fungi spores genuses ?

with colors/ bright photos ( send it to me please )

19 February 2024 7,084 0 View

Trouble cloning of a bacmid miniF replicon. The fragment seems to be ligated but only some part of the fragment is recovered from the plasmid?

Hi, I have been trying to clone 8.4kb miniF replicon from bacmid. I have tried doing that in pFastBAc backbone but the fragment seems to be lost after every trial. While doing colony PCR, I am...

19 February 2024 1,176 1 View

I want to know how to prepare solution that use for autozero and base collection in the uv spectrophotometer?

I have dissolve my extracts using DMSO and I got absorbance that increase with the concentration. I have dough when I prepare measurements in the spectrophotometer I only use 2 sample of DMSO and...

13 February 2024 2,390 0 View

DdPCR Auto dg consumables?

Hello, Is there any alternative (cheaper) Bio-Rad AutoDG-compatible tips? The price for a pack of 20 boxes has become double in 2024. Thanks

13 February 2024 1,312 0 View

Selection of buffers for viability dye and antibody cocktails in flow cytometry?

I am trying to create a workflow for flow cytometry experiment sample prep. I intend to use LIVE/DEAD™ Fixable Aqua Stain and an antibody cocktail for extracellular antigens. I intend to use the...

12 February 2024 6,453 1 View

Does Abaqus subroutine for Crystal Plasticity work?

I watched Youtube tutorials for Abaqus subroutine for crystal plasticity analysis: https://www.youtube.com/watch?v=_0uk_mdyMZY&t=530s https://www.youtube.com/watch?v=D5MJ0D1OMF8 The...

11 February 2024 2,421 3 View

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