2 pts. What is the specific control purpose of PCR tubes 5 and 6 in your PCR pro
ID: 3165833 • Letter: 2
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2 pts. What is the specific control purpose of PCR tubes 5 and 6 in your PCR protocol (transgenic positive control DNA)? 6. 2 pts. Why do we resolve PCR products by gel electrophoresis? 7. 2 pts. What is the purpose of running a molecular mass marker alongside your samples in gel electrophoresis? 8. 6 pts. Pick an Arabidopsis gene of choice (e.g.-FLOWERING LOCUS T. Retrieve its sequence from NCBI's database (https://www.ncbi.nlm.nih.gov/nucleotide). Attach a print out of the sequence to the back of your worksheet. List a pair of primers of amplification of the gene. What size fragment (in bp) would you expect to be amplified with these primers?Explanation / Answer
5. Positive control helps us to check whether the amplified product is similar to the transgenic DNA. It will help us to avoid errors while performing an experiment. So, keeping a positive control (one which gives always positive result) and a negative control (one which does not show any results) is very much essential while performing pcr.
6. Gel electrophoresis is size-dependent separation technique in which DNA is allowed to pass through a gel matrix in presence of charge. Depending on the size, the DNA migrates in the gel and this helps us to determine the exact molecular weight of the DNA. Similarly PCR products are resolved on the agar server gel electrophoresis to determine the molecular weight of the amplified product. It will also help us to remove unwanted primers used in the pcr reaction.
7. Molecular mass markers consist of several DNA fragments of varied molecular weight. After performing electrophoresis, they separate and show a distinct pattern of bands. Each band in this marker has a standard molecular weight and it is used to compare the molecular weight of the amplified product. Hence they are loaded alongside your samples in gel electrophoresis.
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