Can someone help me on how to interpret this gel image of pcr product? Im having trouble in understanding it. These samples were going to sequence. What is the importance of this gel image result before it will undergo dna sequencing? The samples are: A) Serattia, B) Shigella, C) Klebsiella , D) Clostridium, E) Micrococcus, F) Klebsiella, G) Bacillus, H) Neisseria, I) Salmonela, J) Pseudomonas, K) Micrococcus. Thats all. thank you.
Hi Sittie,
The gel image of PCR product will give the product size of your targeted DNA (i.e Bacterial DNA). As you can see, there are different band size in your gel. So, select your target size, purify and send for DNA sequencing!
Good luck!!
Sittie:
Niege Mendes notes important points. Controls help identify and potentially resolve problems. Which band size do you expect? It is important to optimize your PCR reaction conditions including both the reaction mixture (e.g. MgCl2 concentration) and the cycling conditions. Another factor to consider is your sequencing approach. Will you be using internal primers of your target PCR product, sequence of the amplimers (primers), etc? This can have an impact on if "contaminant" or other PCR artifacts affect the sequencing.
Hi Sittie,
As you already got some good response from the three individual researcher, I just wanted to add that you just need to purify your desired PCR product and send it to sequencing but according to you gel picture might be the dominant bands are genomic DNA and your PCR was not working well. So, you should use positive and negative control and then confirm your desired PCR products to send in sequencing!
Bonsoir, augmenter le temps de migration et vous aurez des bandes plus nettes.
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