the nucleotide sequence of the gene encoding for the enzyme luciferase from the firefly

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the nucleotide sequence of the gene encoding for the enzyme luciferase from the firefly (Photinus pyralis) was isolated and sequenced= search about it in general.
Effect of pH on luciferase Activity-Effect of temperature on luciferase activity (search about it in general)
Also, look to the method it will help what kind of information I want.
5 pages for introduction
 
* For the method please make in paragraphs and avoid starting a sentence with a number, using personal pronouns (e.g. ‘I’, ‘we’, ‘our’) and improper units or symbols (e.g. use µL not uL).
at least 12 references of peer-reviewed scientific articles are required.
method
 
*Activity was measured as lumens produced per minute (measured using a spectrophotometer at 450 nm)
 
*The reaction buffer was 100 mM TRIS, 2 mM EDTA, 2 mM MgCl2, 500 mM luminol
*For pH studies, the TRIS buffer was changed from pH 1-14 at 30°C.
*For temperature studies, the pH was 7 and temperature changed from 5-70°C.
 
*For seqeuncing I took 1 g of fireflies, ground the tissue with a mortar and pestle, and extracted genomic DNA using the
“Quick DNA” kit from Bio-Rad.  I used PCR to amplify the luciferase gene using these primers:
LUC-F   5′-ATGGCTGGCCTTGCTACGGGC-3′
LUC-R  5′-AATCTACCATGTTTGGTTGAC-3′
*I then used 22 uL of “Quick PCR Reagent” (Bio-Rad) with 1 uL of genomic DNA and 1 uL of each primer (above).
Thermocycler conditions were: 94°C for 10 min, 35 cycles of [94°C for 30 s, 55°C for 30 s, 72°C for 1 min], followed by
72°C for 10 min.
*I sent the amplified product for sequencing to TCAG Laboratories and processed the sequence using the FinchTV program.
 
*For enzyme analyses, I extracted protein from 1 g of fireflies (with mortar and pestle) using a buffer consisting of 50 mM HEPES (pH 7.0), 2 mM EDTA, 2 mM MgCl2. I filtered the extract through cheesecloth and then centrifuged the suspension at 13,000 rpm for 10 min at 4°C, and then collected the supernatant (containing the enzymes) for the activity analyses.

Our academic experts are ready and waiting to assist with any writing project you may have. From simple essay plans, through to full dissertations, you can guarantee we have a service perfectly matched to your needs.

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ORDER NOW DISCOUNT CODE >>>> WELCOME40

 

 

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the nucleotide sequence of the gene encoding for the enzyme luciferase from the firefly

Our academic experts are ready and waiting to assist with any writing project you may have. From simple essay plans, through to full dissertations, you can guarantee we have a service perfectly matched to your needs.

GET A 40% DISCOUNT ON YOU FIRST ORDER

ORDER NOW DISCOUNT CODE >>>> WELCOME40

 
the nucleotide sequence of the gene encoding for the enzyme luciferase from the firefly (Photinus pyralis) was isolated and sequenced= search about it in general.
Effect of pH on luciferase Activity-Effect of temperature on luciferase activity (search about it in general)
Also, look to the method it will help what kind of information I want.
5 pages for introduction
 
* For the method please make in paragraphs and avoid starting a sentence with a number, using personal pronouns (e.g. ‘I’, ‘we’, ‘our’) and improper units or symbols (e.g. use µL not uL).
at least 12 references of peer-reviewed scientific articles are required.
method
 
*Activity was measured as lumens produced per minute (measured using a spectrophotometer at 450 nm)
 
*The reaction buffer was 100 mM TRIS, 2 mM EDTA, 2 mM MgCl2, 500 mM luminol
*For pH studies, the TRIS buffer was changed from pH 1-14 at 30°C.
*For temperature studies, the pH was 7 and temperature changed from 5-70°C.
 
*For seqeuncing I took 1 g of fireflies, ground the tissue with a mortar and pestle, and extracted genomic DNA using the
“Quick DNA” kit from Bio-Rad.  I used PCR to amplify the luciferase gene using these primers:
LUC-F   5′-ATGGCTGGCCTTGCTACGGGC-3′
LUC-R  5′-AATCTACCATGTTTGGTTGAC-3′
*I then used 22 uL of “Quick PCR Reagent” (Bio-Rad) with 1 uL of genomic DNA and 1 uL of each primer (above).
Thermocycler conditions were: 94°C for 10 min, 35 cycles of [94°C for 30 s, 55°C for 30 s, 72°C for 1 min], followed by
72°C for 10 min.
*I sent the amplified product for sequencing to TCAG Laboratories and processed the sequence using the FinchTV program.
 
*For enzyme analyses, I extracted protein from 1 g of fireflies (with mortar and pestle) using a buffer consisting of 50 mM HEPES (pH 7.0), 2 mM EDTA, 2 mM MgCl2. I filtered the extract through cheesecloth and then centrifuged the suspension at 13,000 rpm for 10 min at 4°C, and then collected the supernatant (containing the enzymes) for the activity analyses.

Our academic experts are ready and waiting to assist with any writing project you may have. From simple essay plans, through to full dissertations, you can guarantee we have a service perfectly matched to your needs.

GET A 40% DISCOUNT ON YOU FIRST ORDER

ORDER NOW DISCOUNT CODE >>>> WELCOME40

 

 

Posted in Uncategorized