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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.