Cloning Brachyury from SW480 in pNEB193 plasmid - Essay Example

Comments (0) Cite this document
Hence, all the cells were removed and stored them in preparation for the RNA extraction. Examinations and recordings were done to examine if the cell count and cell morphology were allowed before the step of…
Download full paperFile format: .doc, available for editing
GRAB THE BEST PAPER94.2% of users find it useful
Cloning Brachyury from SW480 in pNEB193 plasmid
Read TextPreview

Extract of sample
"Cloning Brachyury from SW480 in pNEB193 plasmid"

Download file to see previous pages igure 2: The total RNA was extracted from SW480 cells by use of Norgen’s Total RNA Purification Kit, the samples were then denatured in rapid Formalin – free RNA loading buffer which had Formalin – Free RNA dye. They were then incubated for 5 minutes at a temperature of 700C. Lane number one was filled with RNA ladder 5 μL. The lanes from number 2 to 20 contained 10 μL of each of the class samples. The image capture was then done using GelDocEZ system.
3-Table 1: The concentration and purity of the total extracted RNA from the SW480 cells for the sample H was is shown in table 1. The resulting concentration of RNA was 82.88 ng/ul. An RQI of 7.3 indicated that the RNA quality was accepted. The ratio (28S/18S) was 0.93 though the recommended ratio is 2
The plasmid preparation experiment was undertaken before the start of the RNA extraction. The purpose of this experiment was to purify enough linearized phosphatise treated pNEB183 plasmid to be utilised in the ligation reaction. This purification was attained through several steps that started with the purification of the inoculated plasmid from E.coli culture in LB/ampilicin broth. Using Qubit analysis, the concentration of the purified plasmid was calculated to be 5.1 ul. The EcoR1 enzyme was then utilised to digest the circular plasmid into a linear plasmid which was then treated using alkaline phosphatise enzyme to remove the 5’ phosphate group and hinder self – ligation. The sample was then loaded on 0.8% agarose gel so as to visualize and purify the linearized plasmid from the gel by use of purification method as shown in figure 5:
In figure 5, two bands were clearly seen in the second lane. The first band was an uncut plasmid. The second band was a linear plasmid. Adequate preparation took place as the linear plasmid could migrate longer on the gel. The second band was bright and shiny. It was 2797 bps in length. It also contained ...ul/mg concentration of plasmid. Using x tracta Gel Extraction ...Download file to see next pagesRead More
Cite this document
  • APA
  • MLA
(“Cloning Brachyury from SW480 in pNEB193 plasmid Essay - 3”, n.d.)
Cloning Brachyury from SW480 in pNEB193 plasmid Essay - 3. Retrieved from https://studentshare.org/biology/1635684-cloning-brachyury-from-sw480-in-pneb193-plasmid
(Cloning Brachyury from SW480 in pNEB193 Plasmid Essay - 3)
Cloning Brachyury from SW480 in pNEB193 Plasmid Essay - 3. https://studentshare.org/biology/1635684-cloning-brachyury-from-sw480-in-pneb193-plasmid.
“Cloning Brachyury from SW480 in pNEB193 Plasmid Essay - 3”, n.d. https://studentshare.org/biology/1635684-cloning-brachyury-from-sw480-in-pneb193-plasmid.
  • Cited: 0 times
Comments (0)
Click to create a comment or rate a document
Molecular biology cloning and Brachyury gene
Cloning entails the intensification of specific DNA segments, leading to the buildup of large amounts of similar DNA fragments, which may be utilized for numerous functions. Conventional molecular cloning uses tailored DNA cloning vectors, which are usually derived from spherical DNA molecules identified as bacterial plasmids.
4 Pages(1000 words)Essay
Plasmid mapping
They replicate independently and use a variety of mechanisms to maintain their copy number. They contain the gene codes for the enzymes that are important for the bacterial hosts. The plasmids act as vectors in the molecular biology experiments. The vectors are the carrier DNA molecules into which the foreign gene of interest is inserted and expressed in the host.
7 Pages(1750 words)Essay
Cloning cdc2
Tables have also been shown to represent the results either of the concentrations of the DNA of the plasmid and the genomic DNA concentration. Other results that have been tabulated are the transformation data for both the blue and white colonies. The
8 Pages(2000 words)Essay
However at the same time, the majority of us are in fact not completely sure as to what cloning actually is. In order to come to a clearer and more understanding viewpoint on this subject matter, the elements surrounding the topic of cloning must be properly addresses.
4 Pages(1000 words)Essay
trol, genetic engineering, transplanted heads, computer poetry, and the unrestrained growth of plastic flowers." (Notes of a biology watcher: on cloning a human being. N Engl J Med. 1974; 291:1296-97). It is this statement that serves as the basis of this paper. It is apparent
3 Pages(750 words)Essay
Cloning by artificial means is the one most popularly identified with cloning: It is the creation of a clone scientifically, that is, by some conscious design or human intervention. Contrary to widespread belief,
6 Pages(1500 words)Essay
However, the cloning of plants has been present for longer. The process involves the generation of genetically identical cells from one ancestral cell. For plants it is mainly utilized to create species with the ideal
2 Pages(500 words)Essay
Cloning Brachyury from SW480 in pNEB193 plasmid
2 – The starting point of the practical was the purification of RNA from SW480 human cells. The sample (F) which was mine had a concentration of 308 ng/ μL. This was the
4 Pages(1000 words)Essay
Brachyury gene
hare an exclusive DNA binding domain, and are responsible for the regulation of a myriad of early vertebrate developmental processes (Santa Cruz Biotechnology, Inc. 2013, par. 2), including oogenesis, neurogenesis, the maturity of limb all through the kingdom animalia, and
1 Pages(250 words)Essay
Though the issue of cloning human beings has received criticisms from different sectors of society, it can be observed that some people advocate for this process as a result of different reasons. For instance, parents
2 Pages(500 words)Essay
sponsored ads
We use cookies to create the best experience for you. Keep on browsing if you are OK with that, or find out how to manage cookies.
Let us find you another Essay on topic Cloning Brachyury from SW480 in pNEB193 plasmid for FREE!
logo footer
Contact us:
Contact Us Now
FREE Mobile Apps:
  • StudentShare App Store
  • StudentShare Google play
  • About StudentShare
  • Testimonials
  • FAQ
  • Blog
  • Free Essays
  • New Essays
  • Essays
  • Miscellaneous
  • The Newest Essay Topics
  • Index samples by all dates
Join us:
Contact Us