write a primer
Description
design two new primers (one forward and one reverse) to amplify the GFP gene from the plasmid pGFPuv. A document with an example can be found below.
Important notes:
You must be able to digest the PCR product with the restriction enzyme PstI, so the palindrome must be in your primers or in the sequence between the primers.
Your primers should be between 15 and 25 base pairs and should only appear once in the pGFP sequence.
The Tm of both primers should be shown. Please note, that restriction enzyme sites added onto the primer do not contribute to their Tm.
- Both primers should be shown aligned against the amplified pGFPuv sequence and all elements should be labelled (refer to the example provided).
Unformatted Attachment Preview
Name tudent # – Group #
Forward primer
5 ACGCCAAGCTTGCATGCCTGCAGGT Ò
5`ACGCCAAGCTTGCATGCCTGCAGGTCGACTCTAGAGGATCCCCGGGTACCGGTAGAAAAAATGAG
TAAAGGAGAAGAACTTTTCACTGGAGTTGTCCCAATTCTTGTTGAATTAGATGGTGATGTTAATG
GGCACAAATTTTCTGTCAGTGGAGAGGGTGAAGGTGATGCAACATACGGAAAACTTACCCTTAAA
TTTATTTGCACTACTGGAAAACTACCTGTTCCATGGCCAACACTTGTCACTACTTTCTCTTATGG
TGTTCAATGCTTTTCCCGTTATCCGGATCATATGAAACGGCATGACTTTTTCAAGAGTGCCATGC
CCGAAGGTTATGTACAGGAACGCACTATATCTTTCAAAGATGACGGGAACTACAAGACGCGTGCT
GAAGTCAAGTTTGAAGGTGATACCCTTGTTAATCGTATCGAGTTAAAAGGTATTGATTTTAAAGA
AGATGGAAACATTCTCGGACACAAACTCGAGTACAACTATAACTCACACAATGTATACATCACGG
CAGACAAACAAAAGAATGGAATCAAAGCTAACTTCAAAATTCGCCACAACATTGAAGATGGATCC
GTTCAACTAGCAGACCATTATCAACAAAATACTCCAATTGGCGATGGCCCTGTCCTTTTACCAGA
CAACCATTACCTGTCGACACAATCTGCCCTTTCGAAAGATCCCAACGAAAAGCGTGACCACATGG
TCCTTCTTGAGTTTGTAACTGCTGCTGGGATTACACATGGCATGGATGAGCTCTACAAATAATGA
ATTCCAACTGAGCGCCGGTCGCTACCATTAC -3³ CGCGGCCAGCGATGGTAATGGACGTCATTA 5everse primer
Green
Red
=
=
gene for GFP
PstI restriction enzyme site
Tm forward primer
Tm reverse primer
=
=
80 Ê66 Ê
Purchase answer to see full
attachment
Forward primer
5 ACGCCAAGCTTGCATGCCTGCAGGT Ò
5`ACGCCAAGCTTGCATGCCTGCAGGTCGACTCTAGAGGATCCCCGGGTACCGGTAGAAAAAATGAG
TAAAGGAGAAGAACTTTTCACTGGAGTTGTCCCAATTCTTGTTGAATTAGATGGTGATGTTAATG
GGCACAAATTTTCTGTCAGTGGAGAGGGTGAAGGTGATGCAACATACGGAAAACTTACCCTTAAA
TTTATTTGCACTACTGGAAAACTACCTGTTCCATGGCCAACACTTGTCACTACTTTCTCTTATGG
TGTTCAATGCTTTTCCCGTTATCCGGATCATATGAAACGGCATGACTTTTTCAAGAGTGCCATGC
CCGAAGGTTATGTACAGGAACGCACTATATCTTTCAAAGATGACGGGAACTACAAGACGCGTGCT
GAAGTCAAGTTTGAAGGTGATACCCTTGTTAATCGTATCGAGTTAAAAGGTATTGATTTTAAAGA
AGATGGAAACATTCTCGGACACAAACTCGAGTACAACTATAACTCACACAATGTATACATCACGG
CAGACAAACAAAAGAATGGAATCAAAGCTAACTTCAAAATTCGCCACAACATTGAAGATGGATCC
GTTCAACTAGCAGACCATTATCAACAAAATACTCCAATTGGCGATGGCCCTGTCCTTTTACCAGA
CAACCATTACCTGTCGACACAATCTGCCCTTTCGAAAGATCCCAACGAAAAGCGTGACCACATGG
TCCTTCTTGAGTTTGTAACTGCTGCTGGGATTACACATGGCATGGATGAGCTCTACAAATAATGA
ATTCCAACTGAGCGCCGGTCGCTACCATTAC -3³ CGCGGCCAGCGATGGTAATGGACGTCATTA 5everse primer
Green
Red
=
=
gene for GFP
PstI restriction enzyme site
Tm forward primer
Tm reverse primer
=
=
80 Ê66 Ê
Purchase answer to see full
attachment
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