I am using LB/circle grow to grow bacterial cells and isolate plasmid dna. I am looking for an easy to prepare in-house media. Can any one suggest?

Kesav Kumar Bheemavarapu @Kesav_Bheemavarapu

07 July 2018 0 4K Report

Though I am using either LB broth or circle grow media to grow bacterial cells, I am looking for high cell density easy to prepare media rather than procuring commercially. Can any one suggest me.

Badges
Science topic

More Kesav Kumar Bheemavarapu's questions See All

Is there any method to detect mutations caused by Taq Polymerase with less proofreading capacity and low fidelity?

I have used a Taq Polymerase with less Proof reading and fidelity. I have a doubt that some mutation may be occurred during the PCR. One drawback is the polymerase doesn't have 3'-5' exonuclease...

04 May 2015 8,494 4 View

I am facing problems when I clone my DNA in freedom p CHO vector either at the AVR II/BST site or the ECO RV/ PAC I site. Can any one help me?

Hi all, I am working on freedom p CHO vector. There are two restriction sites called AVR II/ Bst Z171 and ECO RV/Pac I. I am able to digest my vector at the sites and I am able to clone at any one...

02 March 2015 1,986 4 View

Can anyone guide me in learning next generation sequencing software?

Is Biolinux enough?

03 April 2014 9,040 2 View

How to run the BEAST software?

Can anybody explain how to run the BEAST software for bayesian analysis of molecular sequences. Thank you.

08 September 2013 6,699 1 View

How to isoalate DNA from fecal samples

It is very difficult to isolate DNA from fecal samples, can you share the problems and its troubleshooting.

10 November 2011 3,611 3 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