How can obtain E.coli chromosome to insert plasmid?

More Qassim Kshash's questions See All

Please ,any suggestion about increse RBC fragility after exposure lab animals into artificial light 24 hrs?

Lab experiment

02 March 2018 8,527 8 View

What are the specific immunological marker available as kit in lab animals ?

lab expermint

02 March 2018 4,836 7 View

What is the standard pH in the oral cavity that does not allow tooth decay?

Tooth Decay

31 December 2017 3,093 18 View

If the success rate for your students is high or vice versa, does that reflect?

If the success rate for your students is high or vice versa, does that reflect 1- The intellectual maturity of your students 2 - your teaching competence 3 - Ease of questions 4 - students love...

31 December 2017 4,747 20 View

Do you think the level of education is developing or declining in your country?

Education as general

10 November 2017 2,108 22 View

What are the role of universities and graduates in the third world countries ?

In the third world countries there are universities and colleges and a huge number of graduates in all disciplines and there are raw materials and human resources, to see any impact of...

10 November 2017 2,902 13 View

How we can do brain stimulation in depression case?

Brain

09 October 2017 7,946 9 View

What is about the hard-RAT AVAT?

I WANT TO LEARN ABOUT THAT

05 June 2017 1,222 6 View

Which lab animals is prefer to prepare antibodies?

Antibody

04 May 2017 4,569 6 View

Which samples(serum, urine, aqueous humor, CSF) are prefered in PCR detection of leptospira?

Leptospirosis

04 May 2017 171 17 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