BIOTECH LAB FINAL TEST WITH COLLECT ANSWERS 2026 #9
Design a primer to amplify the sequence with only one strand given: 5'
AACTGTAA............CCGTACAT 3' - correct answer 5' AACTGTAA...........CCGTACAT 3'
<<<<<<GGCATGTA 5'
5' AACTGTAA>>>>>>
3' TTGACATT.............GGCATGTA 5'
Lots of AT in primer - correct answer not a good primer
Lots of GC in primer - correct answer good primer
Need to know sequence before designing primers. - correct answer
RUBISCO experiment - correct answer Plant Tissue
V
V
DNA Isolation
V
V
PCR to amplify fragment (rubisco gene) > primers used are universal which means
every rubisco gene is amplified
V
V
Gel electrophoresis > lets you see if PCR is successful
V
V
Cut out band from gel
V
V
Gene clean > clean out fragment from gel > get gene of interest
V
V
Ligate purified pieces with vectors > pgem-T
V
V
Transform in E. Coli using heat shock method
V
V
Select for white colonies > gene inserted in Lac Z genes
V
V
Mini prep > isolate DNA plasmid
V
V
PCR > used SP6/T7 primers
, V
V
Gel electophoresis
V
V
Should be able to see the RUBISCO band at the same position from beginning of
experiment
Why use SP6/T7 primers? - correct answer SP6 > forward primer
T7 > reverse primer
For when we are synthesizing oligonucleotidesthe pgem®-T and pgem®-T Easy
Vectors are high-copy-number vectors containing T7 and SP6 RNA polymerase
promoters anking a multiple cloning region within the α-peptide coding region of the
enzyme β-galactosidase. Insertional inactivation of the α-peptide allows identi cation of
recombinants by blue/white screening on indicator plates.
Nanodrop - correct answer measure concentration of DNA and protein > measures
absorbance
Calculate concentration of milk in samples - correct answer A595 >>> concentration in
mg/ml
Concentration in mg/ml * 50 = protein concentration
Xx g/ cup = 250 ml
Xxxx mg/250 ml = label concentration
Example:
8 g/cup = 250ml
8000mg/250= 32 mg/ml
What is the reporter gene? - correct answer Lac Z gene
AE Buffer - correct answer elution
AW2 buffer - correct answer wash
What is inside GO TAQ Mix? - correct answer 2 dyes- yellow and blue
Reaction buffers
Taq DNA polymerase
Dntps
Mgcl2
PCR profile - correct answer 94 degrees C -denature
54 degrees C - annealing
72 degrees C- extension
PCR product - correct answer amplified rbc-L gene
Design a primer to amplify the sequence with only one strand given: 5'
AACTGTAA............CCGTACAT 3' - correct answer 5' AACTGTAA...........CCGTACAT 3'
<<<<<<GGCATGTA 5'
5' AACTGTAA>>>>>>
3' TTGACATT.............GGCATGTA 5'
Lots of AT in primer - correct answer not a good primer
Lots of GC in primer - correct answer good primer
Need to know sequence before designing primers. - correct answer
RUBISCO experiment - correct answer Plant Tissue
V
V
DNA Isolation
V
V
PCR to amplify fragment (rubisco gene) > primers used are universal which means
every rubisco gene is amplified
V
V
Gel electrophoresis > lets you see if PCR is successful
V
V
Cut out band from gel
V
V
Gene clean > clean out fragment from gel > get gene of interest
V
V
Ligate purified pieces with vectors > pgem-T
V
V
Transform in E. Coli using heat shock method
V
V
Select for white colonies > gene inserted in Lac Z genes
V
V
Mini prep > isolate DNA plasmid
V
V
PCR > used SP6/T7 primers
, V
V
Gel electophoresis
V
V
Should be able to see the RUBISCO band at the same position from beginning of
experiment
Why use SP6/T7 primers? - correct answer SP6 > forward primer
T7 > reverse primer
For when we are synthesizing oligonucleotidesthe pgem®-T and pgem®-T Easy
Vectors are high-copy-number vectors containing T7 and SP6 RNA polymerase
promoters anking a multiple cloning region within the α-peptide coding region of the
enzyme β-galactosidase. Insertional inactivation of the α-peptide allows identi cation of
recombinants by blue/white screening on indicator plates.
Nanodrop - correct answer measure concentration of DNA and protein > measures
absorbance
Calculate concentration of milk in samples - correct answer A595 >>> concentration in
mg/ml
Concentration in mg/ml * 50 = protein concentration
Xx g/ cup = 250 ml
Xxxx mg/250 ml = label concentration
Example:
8 g/cup = 250ml
8000mg/250= 32 mg/ml
What is the reporter gene? - correct answer Lac Z gene
AE Buffer - correct answer elution
AW2 buffer - correct answer wash
What is inside GO TAQ Mix? - correct answer 2 dyes- yellow and blue
Reaction buffers
Taq DNA polymerase
Dntps
Mgcl2
PCR profile - correct answer 94 degrees C -denature
54 degrees C - annealing
72 degrees C- extension
PCR product - correct answer amplified rbc-L gene
