Sanger Pr Synthesis Lab

Purpose:

There are many types of PCR. In this lab, we are using normal PCR. This method is different form the Sanger PCR because normal PCR is a procedure that involves DNA template to be amplified by PCR. The difference is the amplification in sequencing. Normal PCR is making copies in a specific region of DNA. The copies is exponential. It copies the template DNA by each cycle. Thus, it is a new strand DNA copied per cycle. Also, it uses reverse and forward primers to copy the template DNA. As the forward copy increases the complimentary region of the DNA while the reverse is trying to lengthen the duplication of our forward primer. Meanwhile, the Sanger PCR is when we use one primer instead of reverse and forward primers. It only copy one strand rather than the reverse strand of the DNA. Therefore, in Sanger PCR it only copies one strand and can’t be used as a template. In this procedure, the result is linear copies and not exponential. One of the article said that Sanger PCR is a method uses the 4 dideoxynucleotides (ddATP, ddTTP, ddGTP, ddCTP) where there is no free 3' OH (hydroxyl) group. When one of these is

Our primers will be the enzyme that bring out the progression of DNA replication. By having DNA polymerase, we can add nucleotides in our DNA. These primers are made by oligonucleotides for target DNA. One of our primer is SL1 Primer. This primer has a sequence of 5’–CAG TCC AGT TAC GCT GGA GTC–3’. This means we have to achieve 5’ end of the cDNA so it will get out for PCR. This primer means is forward primer. It spliced the SL1to be able to accept in site. It is a primer that is pre-mRNA process. Another primer is SR2 primer. SR2 primer has a sequence of SR2 is: 5’–GGT CAG GTA TGA TTT AAA TGG TCA GT–3’. SR2 is a reverse primer. This primer is design for the 3’ end toward the center will be copied and still producing a strand in that