Hi Zaneta,

I am not sure of what interpretation you need. I am answering this question as if you were unable to determine the presence of amplification or not.

You should have an idea of the expected size of amplification product also to interpret the results. You can find out from the primers and the target.

With that you can directly look for the brightest band in the corresponding sample and find out its size based on DNA ruler you have used. If the size of the product matches with that of expected size, the amplification was successful and you can mark that particular sample as positive.

However, if your query is regarding something else, mention that.

Best wishes,

Murthy.

hello Zaneta,

Some of your pcrs did not work possibly due to poor quality dna, some did not work because you have a primer dimer problem ( PD), some did not work possibly due to the annealing temperature being too high  and some worked very well.

So  specifucally

d1s80 was run too high a voltage so is separating badly. You see where there is no product there is a low molecular weight diffuse smear. This is PD where the primers stick to each other and form a small amplimer which amplifies very well so stops the proper reaction from workinhg,  You often see when the is pd there is no product and where there is product there is no pd.

gel 2 cdh3 right hand side pcr did not work at all. try lowering the annealing temperature or running a temperature gradient pcr to get amplification or maybe your primers are degraded .more work needed but plenty of advice on this website for getting pcr working, On the RHS thepcr is working well . Check the samples that did not work and see if they did not work on the other primer sets. If so then the amount or quality of the dna is poor and nothing will work, Check the OD260/280 ratio and clean up these dna if necessary

gel 3 ADLH

 Samples amplified well when there is no PD and badly  or not at all when there is PD.

You have to get rid of PD and your pcrs will work much better,

The best way of doing this is to design good primers but more practically to use a hot start enzyme for your pcr because usually PD happens when the reaction mix is kept too cool for too long with the enzyme present before starting cycling and the primers stick together and extend so you end up with a product which is short but has perfect homology to your primers so it takes over the reaction because it amplifies more efficiently than your proper product.

Prepare your mix on ice and add the enzyme or magnesium last and immediately put the samples onto a hot block and start cycling. If this does not work use less primer in the pcr or put the samples on a hot bloch and add the enzyme when the samples are hot then start the cycler running the pcr cycles

dear Zaneta;

Cannot interpret your results since there are not expected pcr product size on your amplified genes. but we can tell about what and how are your produced bands in the gel.

on the first gel image top line left handed you had 8 samples as we see 4 of them showed band and 3 of them were showed the primer dimer (it may because high temperature of annealing, lots of cycles, or not extracted DNA samples well and my your primers do not work as well)  

also about the samples were showed the bands you have to know the size of your genes you interested if the expected equal to observed size you get correct results, if not you should change the anealling temperature by useing 3 degree temperature above and 3 bellow of your previous temperature.

on the right handed first image you had 4 samples no one showed tha bands even the second one next to the primer showed the broken fragments due to the high anealing temp. or lots of cycles try to reduce number of cycles and use defferent temperature. we had same results with one of the M.Sc student here we used 25 cycle and defferent temp. were solved the results.

and the belowe lines and the other images you have the same problems try to do some of these suggestion if you would like to:

1. if you have primer dimer try to reduce the amount of primers you used and if you use a few of DNA samples increase it ( about 30ul for RAPD PCR and above 50-100 ul for RFLP and specific primers). 

2. if you have more than one bands, reduce the number of cycles.

3. if you have more than one bands but some of them above your target bands and others below your target bands, use defferent teperatures. used touch down pcr.

4. if you have the dimer from the top to the end of the gel try to descover if any contamination, bad handeled materials specialy storage of DNA is not well. you may have broken fragments.

i am sorry if i am not servce you well

good luck