Which Polymerase should I use for obtaining a gene through PCR which is 3Kb long?

More Shilpi Kumari's questions See All

Does anyone use antinuclear antibody (ANA) and anti-Phospholipid antibody ELISA kit for Non Human Primate serum samples?

I am looking for anti-nuclear antibody (ANA) and anti-Phospholipid antibody ELISA kit for Non Human Primate serum samples. Any suggestions highly appreciated. Thank you.

28 February 2021 6,512 3 View

Treating Intermediate Variable in Linear Regression Analysis in SPSS?

While performing the Linear regression analysis in SPSS version 26.0.0. my intermediate/mediation variable goes to the excluded section. Please share your valuable suggestion "How to restrain the...

25 February 2021 4,591 5 View

How is Initial GDP measured ?

Dear All, I am working on estimating Debt-Growth Threshold but confused about some points. My questions are; - How do we calculate initial GDP Per Capita? Shall we do it in STATA or excel file?...

15 February 2021 2,677 2 View

Why the battery connected on the load side for storing purpose is taking the volatge /current at whatever rating it is given in matlab simulation?

I am designing a DAB DC DC Converter and when I am connecting a battery block in Matlab and fix its specification lets say 350 V and 25 A, when I am giving input as 600V, Battery would charge to...

11 February 2021 3,110 1 View

How do I connect DAB DC DC Converter to a battery in matlab simulation ?

I am trying to model a DAB converter and I need to observe the power transfeered to the load side on varying the phase shift between two bridges, hence I couldnt take a constant load e.g...

09 February 2021 6,489 3 View

Having trouble with qPCR amplification plots. My curve's never seem to reach a plateau. Any ideas?

The parameters of the curve all seem acceptable, typically reactions results are above 0.9852, -3.1 to -3.6 slope, and 90-110% reaction efficiency. The results seem to be what I would expect from...

04 February 2021 1,138 3 View

How can I get rid of the bubbles in the SBS membrane?

I used methylcyclohexane/ ethylcyclohexane / n-butyl acetate mixed solvent to dissolve thermoplastic SBS & terpene resin. The mixture is then poured onto a glass substrate. After solvent...

24 January 2021 8,306 3 View

Can anyone provide me a protocl of Native PAGE with the 30% terminal resolving gel concentraion?

I am separating the AuCNs, which are Au nanoclusters. I used the 3% stacking and 30% resolving polyacrylamide from the reference. Eachtime when the resolving gel is polymerizing, plenty of heat...

22 January 2021 8,522 3 View

After G actin polymerization and labelling I didn't got filaments. Why?

Purified G actin and got 42kda band and an additional band just below it. While performing G actin polymerization and labeling with rhodamine-phalloidin I didn't observe filaments under the...

20 January 2021 3,751 2 View

How can I compute the distribution of dipole moment as a function of the angles between the dipole vector and solid surface through Lammps command?

I want to plot the distribution curve of dipole moment as a function of angles between dipole vector and solid surface in the presence of external electric field. My system is consisting a...

17 January 2021 1,508 4 View

How do I increase the yield of my PCR product?

Hello, We would like to increase the yield of our PCR product. We are running a series of PCR reactions that is targeting ~1.1kb sequence. We begin each reaction with ~400pg of template DNA...

02 March 2021 4,029 3 View

Research on Supply Chain Management in Indian context?

Hi, I am planning to apply for the PhD degree in the Supply Chain Mgt. with specific area of "Cold Storage warehouses" during Pandemics and wars. Where lock downs and shut downs are frequent....

02 March 2021 285 2 View

PACYC PCR not working after several attempts. Would anyone have any suggestions?

So, I have been trying to run a pACYC PCR which will be used later on for a Gibson Assembly. However the PCR is not working. I have already tried gradient PCR and changing extension time; however...

02 March 2021 1,146 2 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 View

How to address this issue in the analysis of Real-Time PCR results?

To dear Researchers, I was analyzing a series of concentration for estimation of Real-Time PCR efficiency. The concentration was 1:10. I used MS-excel to evaluate Slope. The result of slope was -8...

01 March 2021 8,683 4 View

No title

Does anyone have the experience of using Taq Man probes in the QIAGEN Rotar- Gene qPCR machine?

01 March 2021 5,311 1 View

MiRNA qpcr problem?

Dear All, mirna primer showing some problem in the melting curve? any idea why? As attached is the melting curve. The forward sequence is obtained from miRBase and reverse primer is universal.

28 February 2021 5,008 4 View

Runs of nucleotides in Sanger sequencing?

I performed site directed mutagenesis, transformation, and then I sent out plasmids for Sanger sequencing and found out that there is extension of DNA just before the stop codon. I am not sure...

27 February 2021 547 3 View

Can thermal cycler machines be problematic during PCR?

Hi, my question is about the heating of thermal cycler machines and I hope some of you had experienced a similar thing previously. There are two thermal cycler machines in the lab(BioRad) and for...

26 February 2021 4,777 4 View

Calculating volume of DNA required from DNA concentration?

Hi I am a bit confused. They are asking me to find out the volume of DNA required in ul (a total of 30-100 ng for genomic DNA) from the DNA concentration in the nanodrop reading which was 404.8...

26 February 2021 5,029 2 View

Bashiru Garba

For a 3kb DNA fragment, the usual Taq DNA polymerase will be sufficient (we have used it successfully). However, other proofreading polymerases for much longer sizes can be used including GoTaq® Hot Start Polymerase by Promega. Pfu polymerases are rather high-fidelity polymerases where extremely pure DNA is required for downstream application like cloning and mutagenesis

Debmalya Roy

Better to use KOD one (TOYOBO) or KOD plus (TOYOBO).

The amplification rate is 1kb/10 sec for KOD plus, very fast, and has strong 3'→5' exonuclease (proof-reading) activity with high fidelity.