PCR Analysis

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Questions related to PCR Analysis

Which are the upstream and downstream primers for the human beta globin gene in PCR?

i want to find these primers sequences and where is the binding in the gene .  I need the primers for the mutation which causes anemia.

18 January 2016 989 2 View

Bio rad c1000 thermal cycler lid knob problem?

Hi evey one. We  have Bio rad c1000 thermal cycler.  Last time I used C 1000 it was ok, but now it's lid force knob is broken. lid knob is fixed and does not move. what shold I do?

01 January 2016 2,891 0 View

After gene knockout, will incorporation of non-target tissue mRNA make significant error in measuring relative expression level against the wild type?

An SLR1 promoter is a tissue-specific promoter for exocyst genes which is strongly expressed only in stigma and not in the pollen or style. Suppose that we have an RNA knockout gene construct as...

01 January 2016 8,616 0 View

Are there any experimental procedures to confirm the presence of cDNA after reverse transcription before pcr?

is there any experimental procedures to confirm the presence of cDNA after reverse transcription before pcr??

01 January 2016 3,245 13 View

Which are the upstream and downstream primers for the human beta globin gene in PCR?

i want to find these primers sequences and where is the binding in the gene .  I need the primers for the mutation which causes anemia.

01 January 2016 8,440 2 View

What is the reason of secondary bands in low annealing temperatures in gradient PCR?

I am working on a PCR. I did gradient PCR to make sure that the annealing temperature mentioned in the reference study is the best temperature for  annealing. I found there were lots of secondary...

27 December 2015 8,160 3 View

Does entering wrong information in rt pcr machine affects the results?

I wonder if entering a wrong information to rtpcr machine would really affect the results. For instance, wrong concentration or dilution factor when labeling the plate before the run.???

24 December 2015 778 1 View

I am getting more than the two maximum PCR bands expected by allele with the correct size from the polymorphisms studied. What is wrong?

Hello, I am studying MAO-A polymorphisms that consist on a series of repetions of promoter region. Therefore, I expect to obtain up to 5 different PCR bands (291,321,336,351,381bp) depending on...

21 December 2015 2,381 4 View

How to calculate the ratio of PCR master mix and DNA?

Before running PCR what is the ratio of Taq, dNTP, Primers, buffer, water and DNA? is there any method for calculating?

18 December 2015 4,738 3 View

What is the reason of secondary bands in low annealing temperatures in gradient PCR?

I am working on a PCR. I did gradient PCR to make sure that the annealing temperature mentioned in the reference study is the best temperature for  annealing. I found there were lots of secondary...

12 December 2015 7,845 3 View

Does entering wrong information in rt pcr machine affects the results?

I wonder if entering a wrong information to rtpcr machine would really affect the results. For instance, wrong concentration or dilution factor when labeling the plate before the run.???

12 December 2015 3,165 1 View

Any tips for designing primers to amplify lariats (aka lariat introns)?

Dear all, By means of using divergent primers, I have been trying to amplify lariat introns from mouse bone marrow/ primary neuronal cultures and 3T3 cells to study circular RNAs. However, I get...

12 December 2015 1,129 0 View

I am getting more than the two maximum PCR bands expected by allele with the correct size from the polymorphisms studied. What is wrong?

Hello, I am studying MAO-A polymorphisms that consist on a series of repetions of promoter region. Therefore, I expect to obtain up to 5 different PCR bands (291,321,336,351,381bp) depending on...

12 December 2015 6,209 4 View

How to calculate the ratio of PCR master mix and DNA?

Before running PCR what is the ratio of Taq, dNTP, Primers, buffer, water and DNA? is there any method for calculating?

12 December 2015 3,574 3 View

Why is there no house keeping gene band on my PCR ?

Hello,             My PCR shown no band of Actin beta. RNA was extract from same animal but different organ (Lung and brain).The brain shown no band of Actin Beta but the lung did.        Actually...

12 December 2015 4,168 7 View

How can I get rid of negative control giving signals?

I am working on Gut microbiota of wild animals and getting amplification in negative control. Although I used all new reagents initially still I have changed all reagents and water but I am still...

12 December 2015 2,804 7 View

Why is my PCR-RAPD result just showing band from one sample only?

i isolated DNA from animal (gastropod), when i tried to amplify the DNA its only showing band from one sample only. I tried many times with many mixture composition but the only band i can get is...

12 December 2015 3,757 10 View

I used PCR product as the template with the primary primers set for second PCR.What name do I give this process? Is it nested or hemi-nested?

After the first round of PCR to amplify B. coli DNA, there was no detectable band after electrophoresis, I then used the PCR product as the template with the same prime set used in the first...

12 December 2015 8,973 5 View

I run PCR products in a gel, however, out of 10 samples I get 2 missing, primers are validated and RNA integrity is v.good, any idea how to fix it?

I used to run gel before with no problems, but lately I started to get samples missing. I do prepare new TBE buffer everytime I ran a gel, but this seems not to be the problem, since I get other...

12 December 2015 2,271 9 View

Should I use PCR or Western blot?

I want to prove the existence of S100, S100A6, GFAP, MOG and MPZ in SG- and CN-sphere as well as in SG and CN tissue. What is more reliable; PCR or Western blot in this case? What do you think?

12 December 2015 3,969 4 View

Why am I getting non-specific bands on my PCR gels?

Hi! Recently started using a new genotyping kit for PCR and all of a sudden started seeing non-specific bands on the gel. At first I thought contamination, but I've tried all new primers and...

12 December 2015 5,607 3 View

What is the optimum concentration of Genomic DNA and primers to get perfect PCR products?

I want to get sharp bands of PCR products on the gel electrophoresis without any tailings, primer dimers or non-specific products. What is the best concentration of both primers and DNA to get...

12 December 2015 9,680 7 View

Why did I get such a poor gel image?

what could be the possible reason for poor  gel image after running a pcr product. please find the attached image?

12 December 2015 320 4 View

Should I use PCR or Western blot?

I want to prove the existence of S100, S100A6, GFAP, MOG and MPZ in SG- and CN-sphere as well as in SG and CN tissue. What is more reliable; PCR or Western blot in this case? What do you think?

08 December 2015 3,624 5 View

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