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1. Introduction.
Blood is the kind of sample most regularly dissected in the clinical lab. Phlebotomy is a procedure for which a needle is used to draw blood from a vein; usually, to do laboratory tests. The professional who do this procedure is called Phlebotomist. There sources where you can draw blood from, this source are venipuncture and capillary or peripheral blood.
1.1 The purpose of this laboratory experiment is for student to learn how to obtain an adequate blood for laboratory tests by drawing blood from a vein.

2. Methods and Materials
2.1 Venipuncture.
1. Approach the patient, identify yourself, the department you represent, and the procedure you are about to perform., identify the patient and explain the procedure to the patient.
2. Assemble equipment and supplies.
3. Prepare the vacuum blood collection system by attaching the needle to the hub and positioning a tube in the holder.
4. Apply the tourniquet and examine the arm for palpable veins.
5. Palpate the veins.
6. Release the tourniquet, cleanse the chosen site with a 70% alcohol swab. Begin at the puncture site selected and move the alcohol pad outward, in concentric circles (experienced phlebotomists are so quick that they may not release the tourniquet during site preparation).
7. Allow the site to air dry.
8. Reapply the tourniquet, making sure that the ends do not touch the prepared site.
9. Ask patient to clench fist tightly.
10. Position the holder in the palm of your hand between your thumb and index finger. Your palm should be pointing to the left if you are right-handed, and to the right if you are left-handed.
11. Uncap the needle. Inspect the needle for manufacturer’s defects.
12. Anchor the vein selected, using the thumb and index finger.
13. Position the needle in the same direction as the vein selected. Insert the needle, bevel up, at a 15-degree angle. The needle should be inserted in one smooth motion. Only the index finger and thumb should move forward to guide the needle into the vein.
14. Release the vein and push the evacuated tube onto the back of the needle. Be sure to keep the holder stationary. Once the tube has been pushed onto the needle, take your hand off the tube. If the stopper of the tube has been punctured by the back of the needle, and blood is not entering the tube, pushing on the tube will not cause blood to enter it.
15. Allow the tube to fill, when the vacuum has been exhausted, blood will no longer enter the tube.
16. Keeping the holder still, pull the evacuated tube of the back of the needle and replace it with the second tube (if the first tube contained an additive, gently invert it while waiting for the second tube to fill).
17. Once blood begins to enter the second tube, release the tourniquet within one minute of application.
18. Pull the evacuated tube off the needle. Allow it to rest in the holder.
19. Place a piece of gauze or a cotton ball over the puncture site, do not push down on the gauze.
20. Remove the needle from the patient’s arm and immediately apply pressure with the gauze.
21. Activate the needle safety device and dispose of unit.
22. If the last evacuated tube collected contains an additive, invert gently several times to mix the blood with the additive.
23. Inspect puncture site, apply bandage if needed.
24. Label the tubes collected IMMEDIATELY as follows
25. Discard materials in appropriate waste receptacle and disinfect work area.
26. Remove gloves, wash hands and leave patient courteously.

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2.2 Capillary puncture.

1. Approach the patient, identify yourself, the department you represent, and the procedure you are about to perform. Using two unique patient identifiers, confirm the identity of the patient.
2. Check requisition order to ensure proper collection of samples.
3. Assemble all the necessary equipment’s.
4.Assess the patient and determine whether a finger or a heel would be appropriate for use. Heels are preferred for capillary puncture in infants less than 1 year of age.
5.The site of blood collection must be warm to ensure the free flow of blood.
6.Hold the area to be punctured with the thumb and index finger.
7.Clean the area with 70% alcohol pad and allow to air-dry.
8.Use a disposable sterile lancet to puncture the area.
9.Wipe away first drop of blood.
10.Apply gentle pressure to area to obtain a suitable specimen. When the tip of the collection tube touches this drop, blood will flow into the tube by capillary action into the bottom of the tube.
11.When the necessary amount of blood is obtained, clean gauze is used to apply gentle pressure on the puncture site. Bandage the site.
12. Discard used lancets at the sharps container.
13. Remove gloves and discard into biohazard container
14. Wash hands using proper procedure

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1. INTRODUCTION

Restriction endonucleases are enzymes which recognize specific short sequence of DNA and cleave the DNA in both the strands (Alfred,2004). Werner Arber in 1960s predicted that the restriction enzyme act on a DNA sequence, called a recognition site. In 1970s, Hamilton Smith purified a restriction enzyme from Haemophilia influenzae(H.infuenzae) and showed that this enzyme cut DNA at specific sequence. The sequence is usually six base pair long. The cutting of DNA molecule into smaller pieces is known as restriction digestion. Smith showed that, when this recognition site is present in H.infuenzae it does not cut the host cell DNA (Heidi Chial, n.d) . There are four different types of restriction endonucleases. They are Type I restriction endonuclease, Type II restriction endonuclease, Type III restriction endonuclease and Type IV restriction endonuclease (Heidi Chial, n.d).

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Restriction endonucleases were primarily used for obtaining DNA fragments for agarose and polyacrylamide gel electrophoresis. Later, restriction enzymes became the workhorse of molecular biology. DNA mapping uses restriction endonuclease. Mutants were made by deleting one or more fragments of DNA. Early days of sangers DNA sequencing made use of restriction enzymes to cut DNA. The doorway to recombinant DNA technology began with a work done using restriction endonuclease and SV40 DNA. SV40 was converted to a vector for recombinant DNA( Roberts,2005).

Type II restriction endonucleases are the most common restriction enzymes. Type II cut DNA within their recognition sites. The recognition sites are palindromic (R.W.Old,1980). EcoRI is a type II restriction endonuclease. They recognize palindromic sequences that are 4 to 8 base pairs(bp) long and cut DNA within the recognition site in the presence of Mg2+ (Alfred,2001). EcoRI recognizes the sequence GAATTC. EcoRI name originates from the species the enzyme was isolated, Escherichia coli (E.coli), R denotes the strain RY13 and I represents it is the first enzyme to be isolated from Escherichia coli RY13. (R.W.Old,1980).

EcoRI was used to cut lambda(?)DNA and pUC19 DNA. Lambda was discovered by Esther Lederberg in 1951 (Casjens.S,2015). Bacteriophage lambda infects E.coli. This infection increases the rate of genetic recombination as a part of infection cycle (Hillyar.C,2012). When the DNA is isolated from the phage particle, it is in the form of a linear duplex molecule. The DNA is 48.5 kb. Lambda DNA forms a circular structure when inserted into the host cell. This is because of the 5′-projections of 12 nucleotides. These projections form the cos site (R.W.Old,1980). Lambda DNA was used as the ladder to determine the molecular weight of fragmented pUC19 DNA.

pUC19 is a cloning vector. It is small, circular and double-stranded. pUC19 plasmid has multiple cloning site within the coding region of LacZ fragment. Restriction site for EcoRI is present within the 54bp multiple cloning site. pUC19 has only one recognition site for EcoRI (Souza Xavier,2009). It has high copy number. pUC19 is commonly used cloning vector that convey tetracycline(Tet) and Ampicillin(Amp) resistance(New Bio Labs,2018).

Fig:1.1 Restriction sites of pUC19 vector( New Bio Labs,2018)

Gel electrophoresis is a technique ud to separate DNA molecules based on size. Agarose is a plant polysaccharide that is used to make a semi-solid gel for gel electrophoresis of DNA(Malacinski,2005). When electric field is applied, DNA migrates to the positive electrode. Larger DNA fragments migrate with a slower speed. Over several hours, DNA fragments migrate over the gel , forming a ladder of distinct bands. Each band contain several DNA molecules of same length(Alberts,2015).
The aim of the experiment is to examine and distinguish plasmid DNA by restriction endonuclease digestion .

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1. Successful projects are important to Hewlett Packard because that allows them to grow and generate new products, services, and procedures. An increase in revenue means they get capital to work on new projects. Successful projects also increase the reputation of a company which will have a positive impact on their future and allow them to be leaders in the industry. If projects fail, HP risks falling behind against rivals, as well as financial losses.
2. The evaluation team should quantify project contributions rationally, meaning anything that could influence the current marketing performance of the company should be taken into consideration. If the business environment changes, the mission and goals of the company may need to change too, and the evaluation team should adapt to those changes as well. The financial selection is important because its what ensures projects generate adequate return on investment. They also make sure financial success is reflected in the portfolio of projects rather than in the financial contributions of individual projects, the emphasis is on supporting business goals rather than personal agendas.
3. The reason considerable attention is paid to the measures HP uses to evaluate its projects is to support the success of the business. When you prioritize projects efficiently you’re able to get more work done and develop an understanding of the company’s strategy, this results in continued success and aligns with business objectives.
4. The aggregate project plan illustrates the number of projects, their size, history, timing, and where they fall on the scales of innovation for processes and products visually, so its easy to understand. You’re able to compare the types of projects being conducted and different project portfolio proposals. The plan of record records the results of the selection process. You’re able to see the priority and headcount of each project, as well as “out-plan” projects.
5. Out-plan projects should only be reconsidered for inclusion when they have a positive effect on the marketing performance, they also have to fit the current business environment. They also may be placed in the pipeline during a review of the portfolio of projects.
6. I was impressed by how HP carefully selects and invests in projects, they reduced the number of projects authorized which allowed them to be more focused and effective. HP has a high level of project management maturity because of their disciplined project selection process.
7. Additional analysis of non-numeric projects is necessary in order to assess risks successfully. In order to adjust to the company’s needs and evaluate risks effectively, they have to involve qualitative methods of analysis. All of this helps maintain focus and initiate success for the business.

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