Dave Gray's Build-A-Gene Class Notes - Session 1: Difference between revisions
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(New page: In session 1, we amplified three items: # ''Primer + RBS'' - This stretch of DNA contains a "primer" (see below) and a Ribosomal Binding Site. The RBS defines where protein translation b...) |
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The "primer" is a short strand of DNA that the enzymes can build on to replicate the entire strand we want. It is the starting point of the new strand. It contains a combination of nucleotides that will allow it to uniquely bind to the right starting point because the complementary sequence to the primer appears only one place in the DNA. | The "primer" is a short strand of DNA that the enzymes can build on to replicate the entire strand we want. It is the starting point of the new strand. It contains a combination of nucleotides that will allow it to uniquely bind to the right starting point because the complementary sequence to the primer appears only one place in the DNA. | ||
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Revision as of 19:39, 5 August 2013
In session 1, we amplified three items:
- Primer + RBS - This stretch of DNA contains a "primer" (see below) and a Ribosomal Binding Site. The RBS defines where protein translation begins. A ribosome links amino acids together in the order specified bymessenger RNA.
- Terminator + Vector - This sequence will stop the production of the EmGFP protein and provide the transport mechanism to carry the gene into the E. coli bacteria.
- Control - To verify that our technique was sound and that we didn't inadvertently pollute our product with unwanted DNA.
Each of these were mixed with PCR Master Mix and primers and then placed into the PCR machine.
The master mix contains the buffer, enzyme and nucleotides. The buffer is a solution with the correct salt and PH levels to support replication. The enzyme does the work of attaching nucleotides to the DNA template to carry out replication.
The "primer" is a short strand of DNA that the enzymes can build on to replicate the entire strand we want. It is the starting point of the new strand. It contains a combination of nucleotides that will allow it to uniquely bind to the right starting point because the complementary sequence to the primer appears only one place in the DNA.
