Primer sequence for PCR to confirm pFAST Bac Dual plasmid inserted in DH5alpha cells?

More Sivasankar Panickan's questions See All

Thresold line value must be same in different runs in time pcr for calculating viral copies?

During standard curve preparation thresold line value is 560. It should be kept same during running test samples for detecting viral rna copies?

15 August 2023 7,730 0 View

How to perform spatial search in higher dimensions efficiently?

I have a 6D grid of approximation order 10. This means, it has a million points: (x, y, z, Vx, Vy, Vz) = (10, 10, 10, 10, 10, 10). Now, I want to find those points of this grid that lie within a...

10 October 2022 2,003 0 View

Can we use egg adapted influenza virus for serum neutralization test in MDCK cells ?

can we use egg adapted influenza virus for serum neutralization test in MDCK cells?Thanks

29 October 2021 2,421 4 View

Do we need adapt influenza viruses in MDCK cells before conducting serum neutralization test with influenza viruses in MDCK cells ?

Do we need adapt influenza viruses in MDCK cells before conducting serum neutralization test with influenza viruses in MDCK cels?? Thanks

29 October 2021 6,004 3 View

Anybody can give detailed protocol for VNT(virus neutralization) assay for influenza virus?

including, need of virus paasage in MDCK? dilutuion of serum 1:2 or 1:5 ? etc

11 July 2021 6,986 3 View

Gel purification of linearized plasmid, gave low yield why?

I am using Qiagen Gel extraction kit for gel purification of linearized plasmid. The procedure is followed as per the kit. if i load 1ug DNA in agarose gel, i am getting 600ng DNA or less after...

16 October 2020 6,798 4 View

Dear researchers, In RE digestion with Nco I i am getting smearing. Can any one please advice?

well1- 2 log ladder well2- uncut plasmid(3.2kb) well3- cut wit NcoI well4-cut wit NcoI tried with less time of incubation, low amount of enzyme doesnot work

10 September 2020 6,822 3 View

How to find the list of journals with "Your Paper Your Way"?

Dear Professors and Researchers, Can anyone tell me how to find a list of journals with the choice of "Your Paper Your Way"? Thanks in Advance Regards, Siva

03 May 2020 2,611 5 View

Having a marine actinobacterium. Sequenced for 16S and found to have 98.58% similarity with Citricoccus yambaruensis. Can it be a new species?

Sequenced a marine actinobacterium for 16S, assembled and identified in Eztaxon. Obtained 98.58% similarity with Citricoccus yambaruensis. Can it be a new species? Can I go for whole genome...

09 July 2018 3,412 5 View

Any simple and efficient protocol to explore the diversity of PKS, NRPS and relevant gene in the mangrove sediments?

I would like to explore the PKS, NRPS and relevant gene diversity in the metagenomic actinobacterial DNA of the mangrove sediment samples to understand the secondary metabolite production pattern....

16 November 2017 1,751 1 View

How to confirm the site-directed mutagenesis result without performing NGS?

I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...

08 August 2024 4,645 2 View

I can't see the ssDNA band after performing asymmetric PCR. Is there any way to do this?

After performing symmetric PCR, PCR purification was performed. Afterwards, asymmetric PCR was performed using the PCR purification product as a template, but no ssDNA band was confirmed in the...

08 August 2024 1,668 3 View

How much total RNA concentration to be extracted from sorted plasma cells from bone marrow of C57BL/6 mice for RT-PCR ?

i have sorted anti-NP specific plasma cells from bone marrow of C57BL/6 mice at certain times after immunization with variable counts and isolated total RNA using TRIZOL method for RT-PCR using...

05 August 2024 8,835 1 View

Where should the ARS sequence be introduced on a plasmid?

Hi all, I need to introduce an ARS (autonomously replicating sequence) in my plasmid but I'm not sure which position would be the best. Does anyone have any suggestion? A picture of the plasmid...

05 August 2024 1,573 4 View

"A Markov-like Model for Patient Progression"?

A Markov-like Model for Patient Progression" Markov Chain Monte Carlo (MCMC) Markov Chain Monte Carlo (MCMC) is a powerful computational technique used to draw samples from a probability...

05 August 2024 10,079 0 View

Why after performing site directed mutagenesis ,I don't see any colony after transformation?

I want to introduce a point mutation (change in one nucleotide) into my gene of interest (DNA binding domain) I have designed primers as recommended on the Data sheet of the kit : -Both primers...

05 August 2024 9,059 3 View

Anyone having idea about VN primer for miRNA primer design ?

How to design VN primer to attach with universal reverse primer

05 August 2024 2,116 3 View

Why my colony PCR results of my recombinant bacterial not showing any results?

I am performing ligation of the plasmid and a target gene. The steps I have taken are: 1. Double digestion of the plasmid and target gene 2. Ligation of the plasmid with the target gene 3....

05 August 2024 2,570 3 View

Low-yield gel extraction problem?

I am having an issue with my gel image where my PCR product is not appearing very bright on the gel. When I perform gel extraction, the A260/280 purity value is very low. I used the Qiagen gel...

05 August 2024 9,798 3 View

Identify the eight key processes of excellent supply chain management and discuss how each of these processes impacts the end customer?

Read the journal article by Douglas M. Lambert, “The Eight Essential Supply Chain Management Processes,” Supply Chain Management Review, Vol. 8, No. 6 (2004), pp. 18-26

04 August 2024 9,919 4 View

Katie A S Burnette

Use one primer that's in your insert and one from a flanking region in the plasmid. Get the full plasmid map & sequence from the product site.

Right now, your PCR product is nearly 1/2 of the plasmid (~2500 bp), which is way too big for simply screening PCR.

Sivasankar Panickan

It does not have any insert , it is naive plasmid, just to bulk produce plasmid transformed in E coli