Sequence of PTZ57R/T plasmid ?

08 August 2018 0 8K Report

Does anyone has the sequence of PTZ57R/T plasmid? I am planning to send my constructs to sequencing that is why I need the map of PTZ57R/T plasmid. Anyone ?

Badges
Science topic

More Furkan Oflaz's questions See All

How can I introduce restriction site in the middle of my insert?

Does anyone know any kit free protocol for adding restriction site in the middle of insert ?

08 September 2018 3,037 0 View

PCR reaction problem ?

I prepared PCR rxn with phire DNA polymerase and my amplicon is 1.6 kb long. As you can see in gel photo the amplicon is located at 1.5 kb. I was expecting it to locate a bit above. I will use it...

06 July 2018 7,506 0 View

TA cloning problem. Is it possible two or more inserts to ligate with a plasmid in TA clonig ?

I did TA cloning with instaclone PCR cloning kit and I got some weird results. I picked 12 white colonies, isolated them and cut with a single cutter enzyme BamHI (I cut with BamHI to linearize...

06 July 2018 9,413 4 View

Protocol for A-Tailing with Taq Polymerase ?

I am planning to clone 1.3-1.5 kb inserts to plasmid by using TA cloning but I am worrying about error rate of Taq polymerase, that is why firstly I am plannig to do PCR with high fidelity enzyme...

06 July 2018 5,860 0 View

Why I got unexpected larger bands after digestion of my vector (with insert) ?

For my cloning experiment, I first ligated my insert (1.5 kb) with linearized vector (2.9 kb ) that is provided with instaclone PCR cloning kit. After plating, I randomly selected 5 colonies from...

06 July 2018 2,352 10 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

Anyone that can help with Maxquant TMT16plex which is missing?

Hi I am very new to proteomics analysis. I have got raw files from core facility running thermofisher TMT 16plex 4 hours. I cannot find on maxquant the option TMT16Plex there is only up to 11...

25 February 2021 1,675 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