Nested pcr - FAQS.TIPS

More Bushra Javaid's questions See All

Can I use the result from a pilot study to decide the minimum sample size ?

Can I use the result from a pilot study to decide the minimum sample size required to compare the means using independent sample t-test ? Or any other way that can help me to determine the...

29 January 2021 7,677 6 View

Yield of Plasmid is low in Maxipreparation of plasmid for 200ml? What might be the reasons for it?

Hi, I am working with a plasmid which is known to have very low copy number and when I am isolating it by midiprep alkaline lysis method, I am getting good amount of plasmid i.e., 16.56ng/ul....

11 December 2020 1,369 3 View

How to fix the fungus for SEM imaging?

I want to perform the SEM of candida sp. and i want to know that the fixation for the bacteria and fungus are different or same?

10 December 2020 6,169 3 View

What may be the effective strategy to design primers of miRNAs for qRT-PCR?

I designed Primers for miRNAs using mirPrimer algorithm. However when l analysed them by Oligoanalyzer 3.1, it shows Primer-dimer and secondary structure formation. I request your suggestion...

19 September 2020 4,149 3 View

Chemical absorption Co2 removal technologies?

Dear All, Can Any one list down commercially available chemical absorption technologies which can be used for absorption of CO2 from syntehsis gas

17 August 2020 627 2 View

What is the minimum sample size that is acceptable to conduct a survey?

I'm doing a study to evaluate the adherence rate of clinicians to a clinical practice guideline using clinical vignettes (an online survey). The study only includes physicians who are dealing with...

31 July 2020 5,804 4 View

Isolating Total RNA from Paraffin blocks by TRI reagent?

Dear All, I got 2 weeks old paraffin blocks of tumor masses. I want to extract total RNA from them. The available reagents in my lab are: 1) Cell dissociation buffer (GIBCO#13151-014) 2) TRI...

30 March 2020 5,067 5 View

To study antioxidant profile which would be better to analyse ,Serum or Tissue from mice?

I am working on toxicity induced by drugs on invivo model(mice) and few compounds that have antioxidant properties that would be used to reduce this toxicity. The organs I would be looking for...

22 March 2020 4,155 6 View

how to purify and quantitate nanoparticles once synthesized for use in invivo studies?

I am working on nanoparticles and its biological applications . My question is once nanoparticles are synthesized by what methods can they be purified to eliminate contaminants,impurities present...

27 February 2020 2,054 5 View

Difference in type of semiconductor ?

Why there is difference between Halleffect and sensing measurement in the type of semiconductor? Example the first declare p type semiconductor while the second declare n type?

23 February 2020 3,217 6 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

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

Has anyone used the Illumina ITS1 primer sets?

Hi everyone, Illumina provides a list of primers to amplify with high taxonomic coverage the ITS1 region for further fungal sequencing, but I cannot find the exact amount necessary of each...

25 February 2021 6,969 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

Joao Gatica

I believe that the most frequent is to use PCR reaction as such and change the set of primers in a second reaction.

I worked with a variant of nested PCR that consists in about 20 cycles of the first reaction PCR and then use this product as DNA in a second reaction with the same reagents for another 20-25 cycles.

good luck

Vaishali Panwar

can you guys give an idea about nested pcr..i would be glad

Han Zhiqun

U can consult documents

Robert R Martin, Delano James, C AndréLévesque. Impacts of molecular diagnosis technologies on plant disease managemen. Annual Review of Phytopathology 2000,38: 207～ 209.