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Mirabel72
See What 2face Did After His Wife Removes Shoes On Red Carpet
~1.4 mins read

If you think that men are passing through a lot of difficulty in life then you never know that women face more challenges than men. If only you know what ladies go through to look perfect for their spouse. Women will slay in over six inches high heel just to look good. They still manage to smile even with the intense pain that comes with the fashion. That is what happened to the beautiful African queen and wife of the popular artist 2Face Idibia on the red carpet during the 14th Headies awards.

What the people cannot stop talking about what the act of love displayed by Innocent Idibia to his loving wife. His attitude got people emotional that such a man with his status was seen holding his shoes as she walks around barefooted. 2Face is such a meek and humble man. How many men can boldly hold their wife's bag in the public?. Some people will not help their wives like this because they are scared of how people will respond.

2Face with his status proves to his fans the true definition of love and that is, humility is needed in a relationship one must come to the same level with the partner so that the relationship will be a happy one. Annie Idibia is lucky to get married to her friend.

See Annie's photo captions.

 

 

2Face and his wife's fans are delighted with the duo. Add shattered they are saying about the video shared by Annie Macaulay Idibia on Instagram.

 

 

 

 

 

You can watch the video here

 

https://www.instagram.com/p/CLmXtUfgkwx/?igshid=tcaaro4vq3u1

 

 

 

 

See the humble man holding his wife and her shoes as well. Men need to learn from 2Face.

 

From Opera News

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Emily27
Moment A Crippled Beggar Was Exposed By A Goat
~1.2 mins read

Moment a crippled beggar was exposed by a crazy goat

A video of a cripple beggar being disturbed by a goat Surfaced online and it was indeed a blessing in disguise as the goat's act revealed that the beggar is actually a conman.

According to the video,the beggar was seated on the floor going about his usual business of begging a goat suddenly came over and started hitting him.

At first the beggar was reluctant, but when the goat kept disturbing him despite all his attempt to make it go away,he was forced to stand up and walk away.

It got more interesting as the beggar walks away,the goat followed suit.The people who were enjoying the drama also encourage the goat to continue the torment.

If God wants to expose someone, there's actually nothing he can not use to reveal any hidden secret.One can only imagine what entered into the goat.

Click to see video

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Zion20
PCR(POLYMARASE CHAIN REACTION
~7.9 mins read

Introduction

Polymerase chain reaction (PCR) refers to a technique employed widely in the basic and biomedical sciences. PCR is a laboratory technique utilized to amplify specific segments of DNA for a wide range of laboratory and/or clinical applications. Building on the work of Panet and Khorana’s successful amplification of DNA in-vitro, Mullis and coworkers developed PCR in the early 1980s, having been met with a Nobel prize only a decade later. Allowing for more than the billion-fold amplification of specific target regions, it has become instrumental in many applications including the cloning of genes, the diagnosis of infectious diseases, and the screening of prenatal infants for deleterious genetic abnormalities.

Fundamentals

The main components of PCR are a template, primers, free nucleotide bases, and the DNA polymerase enzyme. The DNA template contains the specific region that you wish to amplify, such as the DNA extracted from a piece of hair for example. Primers, or oligonucleotides, are short strands of DNA complementary to the 3' end of each target region. Both a forward and a reverse primer are required, one for each complementary strand of DNA. DNA polymerase is the enzyme that carries out DNA replication. Thermostable analogues of DNA polymerase I, such as Taq polymerase, which was originally found in a bacterium that grows in hot springs, is a common choice due to its resistance to the heating and cooling cycles necessary for PCR.

PCR takes advantage of the complementary base pairing, double-stranded nature, and melting temperature of DNA molecules. This process involves cycling through 3 sequential rounds of temperature dependent reactions: DNA melting (denaturation), annealing and enzyme-driven DNA replication (elongation). Denaturation begins by heating the reaction to about 95 C, disrupting the hydrogen bonds that hold the two strands of template DNA together. Next, the reaction is reduced to around 50 to 65 C, depending on the physicochemical variables of the primers, enabling annealing of complementary base pairs. The primers, which are added to the solution in excess, bind to the beginning of the 3' end of each template strand and prevent re-hybridization of the template strand with itself. Lastly, enzyme-driven DNA replication begins by setting the reaction temperature to the amount which optimizes the activity of DNA polymerase, which is around 75 to 80 C. At this point, DNA polymerase, which needs double-stranded DNA to begin replication, synthesizes a new DNA strand by assembling free-nucleotides in solution in the 3' to 5' direction to produce 2 full sets of complementary strands. The newly synthesized DNA is now identical to the template strand and will be used as such in the progressive PCR cycles.

Given that previously synthesized DNA strands serve as templates, the amplification of DNA using PCR increases at an exponential rate, where the copies of DNA double at the end of each replication step. The exponential replication of the target DNA eventually plateaus around 30 to 40 cycles mainly due to reagent limitation, but can also be due to inhibitors of the polymerase reaction found in the sample, self-annealing of the accumulating product, and accumulation of pyrophosphate molecules.

Real-Time PCR

At its advent, PCR technology was limited to qualitative and or semi-quantitative analysis due to limitations on the ability to quantitate nucleic acids. At that time, to verify if the target gene was amplified successfully, the DNA product was separated by size via agarose gel electrophoresis. Ethidium bromide, a molecule that fluoresces when bound to dsDNA, could give a rough estimate of DNA amount by roughly comparing the brightness of separated bands, but was not sensitive enough for rigorous quantitative analysis.

Improvements in fluorophore development and instrumentation led to thermocyclers that no longer required measurement of only end-product DNA. This process, known as real-time PCR, or quantitative PCR (qPCR), has allowed for the detection of dsDNA during amplification. qPCR thermocyclers are equipped with the ability to excite fluorophores at specific wavelengths, detect their emission with a photodetector, and record the values. The sensitive collection of numerical values during amplification has strongly enhanced quantitative analytical power.

There are two main types of fluorophores used in qPCR: those that bind specifically to a given target sequence and those that do not. The sensitivity of fluorophores has been an important aspect of qPCR development. One of the most effective and widely used non-specific markers, SYBR Green, after binding to the minor groove of dsDNA, exhibits a 1000-fold increase in fluorescence compared to being free in solution. However, if even more specificity is desired, a sequence-specific oligonucleotide, or hybridization probe, can be added, which binds to the target gene at some point in front of the primer (after the 3' end). These hybridization probes contain a reporter molecule at the 5' end and a quencher molecule at the 3' end. The quencher molecule effectively inhibits the reporter from fluorescing while the probe is intact. However, upon contact with DNA polymerase I, the hybridization probe is cleaved, allowing for the fluorescence of the dye.

Reverse-Transcription PCR 

Since its advent, PCR technology has been creatively expanded upon, and reverse-transcription PCR (RT-PCR) is one of the most important advances. Real-time PCR is frequently confused with reverse-transcription PCR, but they are separate techniques. In RT-PCR, the DNA amplified is derived from mRNA by using reverse-transcriptase enzymes, DNA polymerases expressed by RNA-containing retroviruses, to generate a complementary DNA (cDNA) library. Using primers sequences for genes of interest, traditional PCR methods can be used with the cDNA to study the expression of genes qualitatively. Currently, reverse-transcription PCR is commonly used with real-time PCR, which allows one to quantitatively measure the relative change in gene expression across different samples.

Issues of Concern

One disadvantage of PCR technology is that it is extremely sensitive. Trace amounts of RNA or DNA contamination in the sample can produce extremely misleading results. Another disadvantage is that the primers designed for PCR requires sequence data, and therefore can only be used to identify the presence or absence of a known pathogen or gene. Another limitation is that sometimes the primers used for PCR can anneal non-specifically to sequences that are similar, but not identical, to the target gene.

Another potential issue of using PCR is the possibility of primer dimer (PD) formation. PD is a potential by-product and consists of primer molecules that have hybridized to each other due to the strings of complementary bases in the primers. The DNA polymerase amplifies the PD, leading to competition for PCR reagents that could be used to amplify the target sequences.

Clinical Significance

PCR amplification is an indispensable tool with various applications within medicine. Often, it is used to test for the presence of specific alleles, such as in the case of prospective parents screening for genetic carriers, but it can also be used to diagnose the presence of disease directly and for mutations in the developing embryo. For example, the first time PCR was used in this way was for the diagnosis of sickle cell anemia through the detection of a single gene mutation.

Additionally, PCR has greatly revolutionized the diagnostic potential for infectious diseases, as it can be used to rapidly determine the identity of microbes that were traditionally unable to be cultured, or that required weeks for growth. Pathogens routinely detected using PCR include Mycobacterium tuberculosis, human immunodeficiency virus, herpes simplex virus, syphilis, and countless other pathogens. Moreover, qPCR is not only used for testing the qualitative presence of microbes but also to quantify the bacterial, fungal, and viral loads.

The sensitivity of diagnostic tools for mutations to oncogenes and tumor suppression genes has been improved at least 10,000 fold due to PCR, allowing for earlier diagnosis of cancers like leukemia. PCR has also enabled more nuanced and individualized therapies for cancer patients. Additionally, PCR can be used for the tissue typing done that is vital to organ implantation and has even been proposed as a replacement for antibody-based tests for blood type. PCR also has clinical applications in the field of prenatal testing for various genetic diseases and/or clinical pathologies. Samples are obtained either via amniocentesis or chorionic villus sampling.

In forensic medicine, short pieces of repeating, highly polymorphic DNA, coined short tandem repeats (STRs), are amplified and used to compare specific variation within genes to differentiate individuals. Primers specific for the loci of these STRs are used and amplified using PCR. Various loci contain STRs in the human genome, and the statistical power of this technique is enhanced by checking multiple sites.

Questions

To access free multiple choice questions on this topic, click here.

Figure Icon

Figure

1. Steps of traditional PCR process 2. Non-specific fluorophore detection in qPCR 3. Specific hybridization probe detection in qPCR. Contributed by Mousa Ghannam

References

1.
Mullis KB. The unusual origin of the polymerase chain reaction. Sci. Am. 1990 Apr;262(4):56-61, 64-5. [PubMed]
2.
Wittwer CT, Herrmann MG, Moss AA, Rasmussen RP. Continuous fluorescence monitoring of rapid cycle DNA amplification. 1997. BioTechniques. 2013 Jun;54(6):314-20. [PubMed]
3.
Ishmael FT, Stellato C. Principles and applications of polymerase chain reaction: basic science for the practicing physician. Ann. Allergy Asthma Immunol. 2008 Oct;101(4):437-43. [PubMed]
4.
Smith CJ, Osborn AM. Advantages and limitations of quantitative PCR (Q-PCR)-based approaches in microbial ecology. FEMS Microbiol. Ecol. 2009 Jan;67(1):6-20. [PubMed]
5.
Brownie J, Shawcross S, Theaker J, Whitcombe D, Ferrie R, Newton C, Little S. The elimination of primer-dimer accumulation in PCR. Nucleic Acids Res. 1997 Aug 15;25(16):3235-41. [PMC free article] [PubMed]
6.
Saiki RK, Scharf S, Faloona F, Mullis KB, Horn GT, Erlich HA, Arnheim N. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. 1985. Biotechnology. 1992;24:476-80. [PubMed]
7.
Mackay IM. Real-time PCR in the microbiology laboratory. Clin. Microbiol. Infect. 2004 Mar;10(3):190-212. [PubMed]
8.
Muldrew KL. Molecular diagnostics of infectious diseases. Curr. Opin. Pediatr. 2009 Feb;21(1):102-11. [PubMed]
9.
Tamaki K, Jeffreys AJ. Human tandem repeat sequences in forensic DNA typing. Leg Med (Tokyo). 2005 Jul;7(4):244-50. [PubMed]
Copyright Â© 2020, StatPearls Publishing LLC.

This book is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits use, duplication, adaptation, distribution, and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, a link is provided to the Creative Commons license, and any changes made are indicated.

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Frankluiz
Ondo Election Free, Peaceful- Observers
~1.4 mins read
A coalition of domestic accredited observers for last Saturday governorship election in Ondo has described the election as peaceful, free, fair, and credible.
The observers said the conduct of the election was satisfactory and best in recent times.
Speaking at a briefing in Akure, the Ondo state capital, spokesman for the observers, Prince Stafford Bisong, said the voters must be commended for displaying maturity and maintaining peace during the conduct of the election.
Bisong, who said there were reported cases of vote-buying and intimidation of voters in some areas, hailed the Independent National Electoral Commission (INEC) for conducting an acceptable election.
He said “Our observers reported that sensitive and non- sensitive materials arrived early as 7:30 am in approximately 95 percent of the polling units across the state making it a great improvement from the Edo state election. Great work in logistics by INEC.
“In as much as we give kudos to security personnel deployed across the polling units for the efficient response in curbing violent situations, we, however, frown at their compromise in working with party agents to perpetrate vote-buying.
READ ALSO: PSC ranks police conducts in Ondo election ‘above average’
“One wrong that was evident on the day of the election was vote-buying in virtually 85 percent of the polling units across the state, even in the presence of security personnel.
“The problems have become a recurring challenge in virtually every one of our elections and it is a puzzle that will continue to undermine the credibility of our elections and democratic process.
“This is not acceptable and it is a practice that must be stopped for the benefit of outgrowing democracy. We implore Nigerian police and other security to be more professional in their conduct during election duties.
“We appreciate INEC for conducting one of the freest election in Ondo state. We adjudged the election to be free and credible.
“The INEC has declared Governor Rotimi Akeredolu as the winner of the last Saturday election for a second term of four years”.
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Ryan29
Symptoms Of Infertility
~0.9 mins read
The main symptom of infertility is not getting pregnant. There may be no other obvious symptoms. Sometimes, a woman with infertility may have irregular or absent menstrual periods. In some cases, a man with infertility may have some signs of hormonal problems, such as changes in hair growth or sexual function.

Most couples will eventually conceive, with or without treatment.

When to see a doctor
You probably don't need to see a doctor about infertility unless you have been trying regularly to get pregnant for at least one year. Women should talk with a doctor earlier, however, if they:

Are age 35 or older and have been trying to conceive for six months or longer
Are over age 40
Have irregular or absent periods
Have very painful periods
Have known fertility problems
Have been diagnosed with endometriosis or pelvic inflammatory disease
Have had multiple miscarriages
Have undergone treatment for cancer
Men should talk to a doctor if they have:

A low sperm count or other problems with sperm
A history of testicular, prostate or sexual problems
Undergone treatment for cancer
Small testicles or swelling in the scrotum
Others in your family with infertility problems.
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Oxlade789
Wayne Rooney 'in Line To Become Derby Boss' With Phillip Cocu On Brink Of Rams Sack
~1.7 mins read
Wayne Rooney is on the brink of becoming the next Derby County manager with Phillip Cocu on the verge of being sacked.
It follows a shoddy start to the season from the Rams, who are 20th in the Championship.
Club chief Mel Morris had ambitions of his team making a go at promotion this season.
But one win in four has left Derby on the wrong end of the table and sums up a largely tepid spell under Cocu.
The Pride Park outfit ended last season in 10th but finished the campaign with four losses on the bounce.
And that form has carried over into the new season, and Cocu's time could be up. 
According to The Sun, Morris has seen enough and the writing is on the wall for the Dutchman.
And in a shock twist of fate, he is prepared to hand Rooney the reins despite him still being a player for the club.
The 34-year-old, who is club captain, only signed for the Rams in a surprise move in January.
Ironically, it was Rooney who most likely kept Cocu in a job after his last-gasp free-kick gave them the win over Norwich.
But a poor result against Watford will spell the end for the ex-PSV manager.
However, Rooney does not have all his coaching badges from the FA yet - meaning Derby would have to hire a fully-qualified coach alongside him.
The Sun report that this would be former boss John Gregory, who last managed the club in 2003. 
The 66-year-old's most recent venture was in India, where he coached Chennaiyin.
It would not be the first time Morris has turned to inexperience for managers, giving Frank Lampard his first job as a coach.
The Chelsea legend took Derby to the play-off final, where they were beaten by Aston Villa, before moving to the Blues.
Rooney has always insisted he is looking to become a manager, and upon his arrival in Derbyshire earlier this year, he said: “I’ve always been interested in it.
“When Louis van Gaal was manager at United, Ryan Giggs was analysing the opponents and I used to go in there for a few hours after training.”
Rooney has technically already managed when he took charge of the England team for Soccer Aid. 

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