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You have been asked to estimate the content of hemoglobin in a test sample. You decide to perform an assay where the concentration of hemoglobin in the test sample can be measured and compare with a hemoglobin standard curve whose range is across 25 to 800 ug/well. The test sample is treated identically (volume assayed is 10 ul) to the standard hemoglobin and gives an absorbance reading of 0.4. By using the kaleidagraph program to analyse the data, the following equation is produced: y = 0.0005 + 0.0018x Using the above information, answer the following questions:

a) what is the amount of hemoglobin in the test sample added to the well expressed in ug?

b) what is the concentration of the hemoglobin in the test sample expressed in g/L?

c) If the red blood cell count expressed as number of cells/L is 5.5 x 1012, what would be the amount of hemoglobin per red blood cell in the test sample? Express the results in picograms/cell.

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Related questions

Evaluation of PCR product electrophoresed on a 0.8% agarose gelshows non-specific bands. The appropriate modification for the nextPCR reaction is to

a )increase the concentration of Taq polymerase.

b) increase the number of PCR cycles.

c) decrease the template denaturation temperature.

d) reduce the concentration of primers.

In the Sanger sequencing method, chain termination is the resultof

a)misincorporation of dUTP instead of dTTP.

b)incorporation of ddGTP.

c)secondary structures in the template strand.

d)sequencing through repeats.

The purpose of EDTA in a DNA storage buffer is to

a)maintain single strands.

b)bind magnesium

c)activate DNases.

d)dissolve DNA into solution.

The thermal cycler ramp refers to the

a)degree of incline of the machine on the laboratory bench.

b)gradient between denaturation and annealing temperature.

c)programming key to set number of PCR cycles.

d)amount of time required to reach the desired temperature.

Evaluation of a blood sample indicates the presence of a clot.Corrective action to salvage this specimen for analysis would mostlikely require

a)digestion with proteinase K.

b)homogenizing the sample under UV irradiation.

c)removal of the clot with urea.

d)requesting a new sample immediately.

Incorporation of biotin into template DNA by nick-translationrequires

a)DNase.

b)ligase.

c)random hexanucleotides.

d)3 temperatures.

An isolated DNA sample is viscous, difficult to pipet, and willnot resuspend in TE buffer after rehydration at 37C for 24 hours.The most appropriate step is to

a)digest the DNA with a frequent cutting restriction enzyme.

b)re-extract the original sample with higher volumes ofreagents.

c)increase the rehydration temperature to 95C.

d)add more TE buffer.

Excess glycerol in a restriction digestion reaction resultsin

a)enzyme degradation.

b)diminished enzyme activity.

c)increased temperature required for digestion.

d)decreased enzyme buffer pH.

Primer dimers will most likely be a result of PCR when

a)3' and 5' primers are designed as 50% GC.

b)DNA template concentration is very low.

c)enhancing agents are added to the master mix.

d)template DNA is initially denatured completely.

Correct operation of a pH meter requires

a)calculation of the electrode millivolt equivalents.

b)standardization of the meter with a range of buffers.

c)documentation of absorbance and standard deviation.

d)verification of pH readings with calf thymus DNA.

The uracil DNA glycosylase pre-PCR decontamination procedurefailed. The most likely cause of the failure is the

a)enzyme solution was heated prior to PCR.

b)enzyme buffer solution was alkaline pH.

c)N-glycosidic DNA bonds cleaved after enzyme treatment.

d)PCR was performed with dTTP instead of dUTP.

The real-time PCR analysis does not show the characteristicmelting curve for the patient or control samples. The fluorimetryreadings of the PCR products indicate the appropriate concentrationof template was added to the real-time PCR reaction mix. Which ofthe following reagents was likely missing from the PCR master mixto account for this finding?

a)SYBR Green I

b)template DNA

c)DMSO

d)5X Cresol Red

Which of the following conditions favor enhanced specificity ofPCR?

a)decrease in the concentration of dNTPs

b)increase in the sodium chloride concentration

c)increase in the concentration of Taq polymerase

d)decrease in the ramp speed and temperature

Which of the following compounds is an inhibitor of Taqpolymerase?

a)magnesium

b)hemoglobin

c)DMSO

d)betaine

Which of the following compositions of gels would allow for theefficient separation of linear DNA molecules in the range of 100 to300 base pairs in length?

a)0.3% agarose

b)1.0% agarose

c)1.0% polyacrylamide

d)8.0% polyacrylamide

4.92*10^4 cpm of a random primed 32P dCTP (3000 Ci/mmol) labeledoligonucleotide probe were precipitated from a 50 uL standardreaction by TCA and 5.28*10^4 cpm is the total cpm in the sample.What is the percent incorporation of radioactivity for thislabeling reaction?

a)1.86%

b)46.50%

c)93.00%

d)98.40%

The stock solution of HCl is 1M. How much stock solution isrequired to prepare 250 mL of 50 mM HCl?

a)0.2 mL

b)5 mL

c)12.5 mL

d)125 mL

Spectroscopy measurement of a DNA sample isolated by saltextraction technique gave the following absorbance readings: A2600.4032 A280 0.3765

This sample is:

a)acceptable for molecular testing.

b)contaminated with salt.

c)contaminated with protein.

d)contaminated with high amounts of RNA.

What is the dilution factor if 50 uL of blood is diluted with950 uL of saline?

a)19

b)20

c)40

d)50

Which of the following are used to prepare a 0.9% agarosegel?

a)0.9 g agarose + 100 mL distilled water

b)0.9 g agarose + 100mL TAE

c)9 g agarose + 100mL TBE

d)90 g agarose + 100 mL buffer

The concentration of double-stranded DNA with an optical densityreading of 1.0 is

a)0.5 ug/ul

b)1.0 ug/ul

c)10 ng/ul

d)50 ng/ul

How much Tris base (MW 121.1) would be required to make 4 litersof Tris-EDTA buffer that is 15 mM Tris?

a)0.7266 grams

b)0.1211 grams

c)1.8165 grams

d)7.266 grams

jadeturtle730

i need question number nine please

Case Study Section 2 â glycolysis
Based on âMonday at the Metabolic Clinic, Adventures in Glycolysisâ by Nancy Boury (National Center for Case Study Teaching in Science)

Background:
You have been chosen for a very competitive undergraduate clinical internship at St. Visintainerâs Hospital and Metabolic Clinic working with Dr. Saccharo, an expert in enzyme deficiency disorders related to glycolysis. Before your first day Dr. Saccharo has asked that you review the reactions of glycolysis and the concept of isoenzymes.

Recall that isoenzymes are groups of enzymes that catalyze the same reaction in that they use the same reactants to product the same products. They may have different genes, tissue expression, developmental timing of expression, and affinities for their substrates. As an example, phosphofructokinase-1 has 3 isoenzyme forms: PFK-L is expressed in the liver and the gene is found on chromosome 21; PFK-M is expressed in the muscle and the gene is found on chromosome 12; and PFK-P is expressed in platelets and the gene is found on chromosome 10.

Question 1:
If a person would be deficient in all 3 forms of PFK-1, this person would .
A. Have lower than normal glycogen stores.
B. Have more mitochondrial activity than normal.
C. Be dead.
D. Produce more energy per glucose molecule because glycolysis would be unregulated.

It is now Monday morning and you are ready to start seeing patients with Dr. Saccharo.

Patient 1:
Ann is a teenager and avid golfer who was referred to the clinic after being refused at the blood drive and tiring easily on the high school golf course during practice. Ann is examined and a blood sample was obtained for analysis of a glycolysis related metabolic panel. The results of her panel is below (levels: + = normal, - = decreased; ++= increased):
Blood Metabolic Panel
Blood Glucose +
Glucose-6-phosphate ++
Fructose-6-phosphate ++
Fructose-1,6,bisphosphate ++
Glyceraldehyde-3-phosphate ++
1,3-bisphosphoglycerate ++
Phosphoenolpyruvate ++
Pyruvate -
ATP -
Red Blood Cell Concentration -
Question 2:
Assuming that Dr. Saccharo is correctly assuming that Ann has a defect in glycolysis, what glycolytic enzyme is the most likely to have the defect based on the blood metabolic profile?
A. Hexokinase
B. Phosphofructokinase
C. Triose Phosphate Isomerase
D. Pyruvate Kinase
E. Aldolase

Patient 2:
Marie is a 32-year-old mother of three complaining of fatigue and muscle cramps with exercise. She had always blamed her intolerance to exercise on her sedentary lifestyle. However, she recently joined a gym and after a week of aerobics classes went to her physician, who ordered a series of blood tests. The blood work came back with abnormal results, leading to her muscle biopsy and referral to the metabolic clinic. Marieâs blood was subjected to the same metabolic panel used before with the following results:
Blood Metabolic Panel
Blood Glucose +
Glucose-6-phosphate ++
Fructose-6-phosphate ++
Fructose-1,6,bisphosphate -
Glyceraldehyde-3-phosphate -
1,3-bisphosphoglycerate -
Pyruvate -
ATP -
Red Blood Cell Concentration -

Marieâs muscle biopsy was also analyzed and showed an excess of glycogen.

Question 3:
What is the most likely enzyme deficiency for Marie?
A. Hexokinase
B. Phosphofructokinase
C. Triose Phosphate Isomerase
D. Pyruvate Kinase
E. Aldolase

Question 4:
If you wanted to test the ability of red blood cells to complete glycolysis what compound would you try to detect?
A. Fructose-6-phosphate
B. Aldolase
C. Pyruvate
D. Phosphofructokinase
E. Dihydroxyacetone phosphate

Question 5:
If you used a sample of Marieâs blood for this, what compound could you add to enable these cells to complete glycolysis?
A. Glucose
B. Fructose-1,6-bisphosphate
C. Insulin
D. Glucagon
E. Fructose-6-phosphate

Patient 3:
Leo is a 25-year-old accountant that is not anemic, has perfect teeth, but has experienced two fainting spells recently. One happened after accidentally drinking a non-diet soda; the other after eating ânaturally sweetenedâ fruit salad. His dentist referred him to Dr. Saccharoâs clinic. Dr. Saccharo suspects Leo may have Hereditary Fructose Intolerance (HFI) and has recommended a closely monitored fructose tolerance test.

Hereditary Fructose Intolerance is a recessive disorder where individuals lack a functional copy of one isoform of the enzyme aldolase, specifically Aldolase B (also known as fructose-1-phosphate aldolase). In the liver, fructose is phosphorylated by Fructokinase to fructose-1-phosphate, then is cleaved by Aldolase B to glyceraldehyde and dihydroxyacetone phosphate. Fructose-1-phosphate is also a known allosteric inhibitor of Glycogen Phosphorylase, the primary enzyme responsible for releasing glucose from glycogen.

Question 6:
As compared to a normal individual that consumes fructose, a person with HFI who ingested fructose would have (select all that apply):
A. High levels of blood fructose
B. High levels of blood fructose-1-phosphate
C. High levels of blood glucose
D. Low levels of blood fructose
E. Low levels of blood glucose

That afternoon Leo undergoes the fructose tolerance test with the following results:

Question 7:
Based on the results of the test, what was the effect of giving Leo the intravenous fructose?
A. A rise in blood glucose levels
B. A decrease in blood glucose levels
C. An increase in glycogen stores
D. A rapid drop in blood pressure

However there were complications: Soon after the test Leo orders and drinks a cup of tea, but after half the cup Leo passes out. Dr. Saccharo orders two sequential blood glucose checks. The first was 50 mg/dl and 15 minutes later Leoâs blood glucose was down to 40 mg/dl. With those results Dr. Saccharo injected something into Leoâs IV and within minutes Leo was back to normal.

Question 8:
From these results does it appear that Leo has HFI?
A. Yes
B. No
C. It is unclear from the results

Question 9:
What did Dr. Saccharo inject into Leoâs IV line?
A. Insulin
B. Glucagon
C. Glucose
D. Fructose
E. Phosphofructokinase

Dr. Saccharo puts Leo on a restricted diet and orders follow-up visits.

Question 10:
Which of the following foods/ingredients would Leo NOT be allowed to eat?
A. Artificial sweeteners
B. High starch foods, such as potatoes
C. Sweet tea made with cane sugar
D. Any dairy products

Dr. Saccharo puts Leo on a restricted diet and orders follow-up visits.

Case Study Section 2 â glycolysis
Based on âMonday at the Metabolic Clinic, Adventures in Glycolysisâ by Nancy Boury (National Center for Case Study Teaching in Science)

Question 9:
What did Dr. Saccharo inject into Leoâs IV line?
A. Insulin
B. Glucagon
C. Glucose
D. Fructose
E. Phosphofructokinase