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Answered on 07 May Tuition/MBBS & Medical Tuition

Neha Bhardwaj

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Yeah definitely.. u can check out old question papers and hit the books immediately. Take advice and correct guidance would make u Excell. Every one in their 1st Prof starts one month before only
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Lesson Posted on 13 Mar Tuition/MBBS & Medical Tuition/Biochemistry

Genomic Library

Varsha Chowdhry

I am an qualified teacher having 7 years teaching experience in Biochemistry. All my chapters are covered...

Genomic Library: Genomic library, produced when the complete genome of a particular organism is cleaved into thousands of fragment and all the fragments are cloned by insertion into a cloning vector. The first step in preparing a genomic library is isolation and digestion of the DNA by restriction... read more

Genomic Library:

Genomic library, produced when the complete genome of a particular organism is cleaved into thousands of fragment and all the fragments are cloned by insertion into a cloning vector.

  • The first step in preparing a genomic library is isolation and digestion of the DNA by restriction endonucleases - This process produces fragments of DNA that include the organism’s entire genome.
  • A plasmid, BAC, YAC or bacteriophage vector is digested with the same enzyme.
  • DNA ligase is used to ligate genomic DNA pieces and vector.

In theory, all DNA fragments in the genome will be cloned into a vector:

  • Recombinant vectors are then used to transform bacteria or yeast cells.
  • Each transformed bacterium or yeast cell grows into a colony, or “clone,” of identical.
  • Cells, each cell-bearing the same recombinant plasmid.
  • Consider each clone a “book “in this “library” of DNA fragments.

Making of Gene Libraries:

There are three general ways to produce genomic libraries:

1. Complete digestion with restriction enzyme, cleave at all relevant restriction sites. This has drawback:

  • Genes containing one or more sites for the restriction enzyme will be cloned into two or more pieces.
  • To screen the entire genome, a very large number of clones would have to be examined, because insert DNA size is relative small.

2. Longer DNA fragments can be generated with mechanical sheering (e.g. passage through a syringe needle) rather than restriction enzyme cutting. A disadvantage is the absence of uniform ends, require enzymatic modification before insertion into a cloning vector.

3. Partial digestion with a restriction enzyme is controlled so that it cuts only some of the available sites. Ideally, this results in cloning a population of overlapping fragments representing the entire genome.

i. Partially digested DNA molecules in a certain size range are selected by density gradient centrifugation or agarose gel electrophoresis.

ii. DNA fragments with sticky ends from restriction digestion can be cloned directly.

iii. Genomic sequences are not equally represented in the libraries, because:

  • Regions of DNA with relevant restriction sites very close together or very far apart axe removed at the size selection.
  • Some regions of eukaryotic DNA prevent vector replication in E coli, and so are eliminated from the library.

One disadvantage of creating this type of library for eukaryotic genes:

  • Is that non-protein coding pieces of DNA called introns are cloned in addition to protein coding sequences (exons), because a majority of DNA in any eukaryotic organism consists of introns.
  • Many of the clones in a genomic library will contain non-protein coding pieces of DNA.
  • Another limitation of genomic libraries is that many organisms, including humans, have such a large genome that searching for a gene of interest really is like searching for a needle in a haystack!
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Lesson Posted on 27/12/2017 Tuition/MBBS & Medical Tuition

BPPV Maneuver

Dr.paparaja Subbanna

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Lesson Posted on 27/12/2017 Tuition/MBBS & Medical Tuition Tuition/MBBS & Medical Tuition/Biochemistry

Glycolysis: Easy To Remember

Dr.paparaja Subbanna

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Lesson Posted on 27/12/2017 Tuition/MBBS & Medical Tuition Tuition/MBBS & Medical Tuition/Biochemistry

TCA Cycle

Dr.paparaja Subbanna

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Lesson Posted on 23/12/2017 Tuition/MBBS & Medical Tuition

Clotting Factors

Dr.paparaja Subbanna

Teaching is my hobby not job. I love teaching. Say any topic I can make it easy for students. According...

Number and/or name Function Associated genetic disorders I. (Fibrinogen) Forms clot (fibrin) Congenital Afibrogenmia Familial Renal Amyloidosis II. (Prothrombin) Thrombin or IIa activates I, V, VII, VIII, XI, XIII, Protein C and platelets Thrombophilia ... read more

Number and/or name

Function

Associated genetic disorders

I. (Fibrinogen)

Forms clot (fibrin)

Congenital Afibrogenmia

Familial Renal Amyloidosis

 

II. (Prothrombin)

Thrombin or IIa activates I, V, VII, VIII, XI, XIII, Protein C and platelets

 

Thrombophilia

 

III. Tissue factor

Co-factor of VIIa

 

IV. Calcium

Required for coagulation factors to bind to phospholipid (formerly known as factor IV)

Hypocalcemia is not associated with clotting problem because it has other more serious effects.

V. (Proaccelerin)

Co-factor of X with which it forms the Prothrombinase complex

Activated protein C resistance

 

VI

Unassigned – old name of Factor Va

 

VII. (Stable factor, proconvertin)

Activates IX, X

congenital proconvertin/factor VII deficiency

VIII. (Anti Hemophilic Factor A)

Co-factor of IX with which it forms the TENASE complex

Haemophilia A

IX. (Antihemophilic factor B or Christmas factor)

Activates X: forms TENASE complex with factor VIII

Haemophilia B

X. (Stuart-Prower factor)

Forms Prothrombinase complex with factor V and converts Prothrombin or II into Thrombin or IIa.

Congenital Factor X deficiency

 XI. (Plasma Thromboplastin Antecedent)

 

Activates IX

Haemophilia C

XII. (Hageman factor)

Activates factor XI, VII and prekallikrein

Hereditary Angioedema type III

XIII. (Fibrin-Stabilizing Factor)

Congenital Factor XIIIa/b deficiency

 

XIV. Von Willebrand factor or vWF

Binds to VIII, mediates platelet adhesion

Von Willebrand disease

 

XV. Prekallikrein(Fletcher factor)

Activates XII and prekallikrein; cleaves HMWK

Prekallikrein/Fletcher Factor deficiency

XVI. High Molecular weight Kininogen or HMWK

 (Fitzgerald factor)

Supports reciprocal activation of XII, XI, and prekallikrein

Kininogen deficiency

XVII. Fibronectin

Mediates cell adhesion

Gomerulopathy with fibronectin deposits

XVIII. Antithrombin III

Inhibits IIa, Xa, and other proteases

Antithrombin III deficiency

XIX. Protein C

Inactivates Va and VIIIa

Protein C Deficiency

XX. Protein S

Cofactor for activated protein C

Protein S deficiency

XXI. Protein Z

Mediates thrombin adhesion to phospholipids and stimulates degradation of factor X by ZPI

Protein Z deficiency

XXII. Plasminogen

Converts to plasmin, lyses fibrin and other proteins

Plasminogen deficiency, type I (ligneous conjunctivitis)

XXIII. Alpha 2 antiplasmin

Inhibits plasmin

Antiplasmin deficiency

XXIV. Tissue plasminogen activator or tPA

Activates plasminogen

Thrombophilia

 

XXV. Urokinase

Activates plasminogen

Quebec platelet disorder

XXVI. Plasminogen Activator Inhibitor 1 (PAI1)

Inactivates tPA & urokinase (endothelial PAI)

Plasminogen activator inhibitor-1 deficiency

XXVII. Plasminogen Activator Inhibitor 2 or PAI2

Inactivates tPA & urokinase

 

XXVIII. Cancer procoagulant

Pathological factor X activation

Thrombophilia

 

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Lesson Posted on 20/11/2017 Tuition/MBBS & Medical Tuition

Immunity Flow Chart

Dr.paparaja Subbanna

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Lesson Posted on 21/11/2017 Tuition/MBBS & Medical Tuition

Cardiac Cycle

Dr.paparaja Subbanna

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Lesson Posted on 19/11/2017 Tuition/MBBS & Medical Tuition

Erythropoiesis

Dr.paparaja Subbanna

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Lesson Posted on 24/08/2017 Tuition/Class XI-XII Tuition (PUC)/Biology Tuition/MBBS & Medical Tuition/Biochemistry Tuition/BSc Tuition/Biochemistry +5 IT Courses/Bioinformatics Tuition/BA Tuition/Advanced cell biology CBSE/Class 10/Science/Acids, Bases and Salts CBSE/Class 10/Science/Acids, Salts and Bases IIT JEE/IIT - JEE Advanced/Chemistry/Physical Chemistry/Atomic Structure and Chemical Bonding less

NEET: Tips For Medical Aspirants

Shaista Jabeen

I am a life science scholar, currently pursuing PhD in a well known reputed college for women. I can...

NEET Exam: 10 chapters in Biology 90 questions Concentrate on only ten chapters in biology that covers almost 90 questions. Five chapters from +1 syllabus and five from +2 syllabus. Chapters to concentrate most: Diversity in living world. Structural organization in animals and plants. Cell... read more

NEET Exam:

  • 10 chapters in Biology
  • 90 questions
  • Concentrate on only ten chapters in biology that covers almost 90 questions. Five chapters from +1 syllabus and five from +2 syllabus.

Chapters to concentrate most:

  • Diversity in living world.
  • Structural organization in animals and plants.
  • Cell structure and function.
  • Plant physiology.
  • Human physiology.
  • All these chapters from +1 syllabus.
  • Reproduction.
  • Genetics and evolution.
  • Biology and human welfare.
  • Biotechnology and its applications.
  • Ecology and Environment.
  • These chapters from +2 grade.
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