Reagents & Solutions
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Characterization of TNT® T7 Quick for PCR DNA:
Compatibility with Other Procedures
Transcend™ Non-Radioactive Translation Detection System
To verify this translation system is compatible with non-radioactive detection,
linearized Luciferase T7 Control DNA* was titrated into the TNT®
T7 Quick for PCR DNA system in the presence of Transcend™ Biotinylated Lysine tRNA (Cat.# L5061). The results of this experiment, depicted in Figure 10 demonstrate that both
colorimetric and chemiluminescent non-radioactive detection using Transcend™ tRNA can
detect protein produced from as little as 10ng luciferase template DNA. As luciferase contains numerous lysine residues,
proteins with fewer lysine residues will possibly exhibit less sensitive detection when
using these non-radioactive detection methods.
Canine Pancreatic Microsomal Membranes (CMMs)
The compatibility of Canine
Pancreatic Microsomal Membranes* (Cat.# Y4041)
with the TNT® T7 Quick for PCR DNA system was investigated
using the b-lactamase and a-mating
factor Control RNAs, which are included with the CMMs (Cat.# Y4061, Y4071). Both RNAs were expressed in this system and
processed as expected (Figure 11). The 31kDa b-lactamase
protein undergoes signal peptide cleavage to 28kDa, while the 18kDa a-mating
factor protein is N-glycosylated to 30kDa.
*Products may be covered by pending or issued patents. Please visit
our patent and trademark web page for more information.
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Figure
10. Titration of linearized luciferase plasmid DNA into TNT® T7 Quick for PCR DNA
using Transcend™ Biotinylated Lysine tRNA incorporation. |
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Figure
11. Expression and processing of b-lactamase and a-mating factor RNAs in TNT® T7 Quick for PCR DNA in the presence or
absence of CMMs. |
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