Worksheet 1: Attempt review
My LMS Subjects / 2021-PSB-MED3LAB(SP-T2-FT) / Assessment / Worksheet 1
Started on Friday, 4 June 2021, 6:00 PM
State Finished
Completed on Sunday, 6 June 2021, 5:33 PM
Time taken 1 day 23 hours
Grade 12.00 out of 15.00 (80%)
Information
Worksheet 1
One of your...
My LMS Subjects / 2021-PSB-MED3LAB(SP-T2-FT) / Assessment / Worksheet 1
Started on
Friday, 4 June 2021, 6:00 PM
State
Finished
Completed on
Sunday, 6 June 2021, 5:33 PM
Time taken
1 day 23 hours
Grade
12.00 out of 15.00 (80%)
Information
Worksheet 1
One of your first tasks in the practicals is to perform a PCR to obtain large quantities of the GFP cDNA from the pE-GFP
vector. Following digestion with appropriate restriction enzymes, this DNA will be ligated in the correct reading frame
into the pQE30 vector, resulting in the production of a GFP-hexahistidine tagged gene fusion. In order to understand this
process fully, it is essential for you to gain some experience in identifying ORFs and restriction enzyme sites. It is also
important that you understand some of the factors involved when designing oligonucleotides to act as PCR primers.
These considerations will be addressed in this activity.
The activity is divided into 3 Parts and are designed to complete in order. For each part, you will be required to download a
PowerPoint slide and use the instructions on the slide to complete the exercises. After you complete each section, you
should answer the associated questions within the quiz.
Note: As this is an interactive PowerPoint slide, it is essential that you do not change the sizes of text boxes or
font sizes when working through each activity.
PART A: THE pQE30 VECTOR
This activity should be used for examining the features of the pQE30 vector and establishing what the vector will
theoretically look like following digestion with restriction enzymes.
To get started, download Part A of the activity here.
You should save a copy of this PowerPoint file onto your own computer and work through the activity. Once
complete, you can answer the associated quiz questions and upload your activity below.
Question 1
Correct
Mark 2.00 out of 2.00
Within the pQE30 vector sequence, you should have identified the region encoding the 6xHis-tag.
Write the DNA nucleotide sequence that encodes for this tag here:
CATCACCATCACCATCAC
NB. You should include the sequence for the 'coding' strand only (5' →3'). Use capital letters to enter your response. Do not
include numbers or spaces as a part of your answer.
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