Producing Fragments

Reverse Transcriptase

The target gene for producing recombinant DNA is obtained by producing DNA fragments. DNA fragments can be produced using reverse transcriptase.

mRNA

mRNA is a single-stranded molecule that is produced when a specific length of DNA (the target gene) is transcribed.
mRNA is complementary to the base sequence in the target gene.
The mRNA sequence can be used as a template for producing fragments of DNA.

Reverse transcriptase

Reverse transcriptase is an enzyme that converts single-stranded mRNA into double-stranded DNA.
Reverse transcriptase can be used in producing DNA fragments by converting the mRNA for the target gene into double-stranded DNA.
The DNA produced from reverse transcriptase is called complementary DNA (cDNA).

E.g. Insulin

DNA fragments of insulin are isolated from pancreatic cells in the following steps:
- mRNA for insulin is isolated from the pancreatic cells.
- The mRNA is mixed with reverse transcriptase.
- Reverse transcriptase converts mRNA into cDNA.
- The cDNA can now be used to produce recombinant DNA.

Restriction Endonucleases

The target gene for producing recombinant DNA is obtained by producing DNA fragments. DNA fragments can be produced using restriction endonuclease enzymes.

Recognition sequences

Recognition sequences are sections of DNA where the base sequence has palindromic base pairs.
- Palindromic base pairs have a sequence of base pairs that are the same but in opposite directions.
Recognition sequences can be used to isolate the target gene if there are two sets of sequences either side of the gene.

Restriction endonucleases

Enzymes called restriction endonucleases bind to recognition sequences.
Each restriction endonuclease binds to a specific recognition sequence (e.g. Eco RI is a restriction endonuclease that binds to GAATTC).
If two restriction endonucleases bind to two recognition sequences surrounding a target gene, the target gene can be cut out of the DNA.

Producing the fragment

DNA fragments can be produced in this way using restriction endonucleases. The steps involved are -
- DNA containing the target gene is mixed with the restriction endonucleases.
- Restriction endonucleases bind to the recognition sequences on either side of the target gene.
- The target gene is cut out of the DNA.

Gene Machines

The target gene for producing recombinant DNA is obtained by producing DNA fragments. DNA fragments can be produced using gene machines.

Synthesising DNA

DNA fragments can be produced by synthesising the target gene sequence using free-floating nucleotides.
This is useful because a DNA template isn't necessary.
The sequence for the target gene is obtained from a database.

Producing the fragment

DNA fragments can be produced in this way in the following steps -
- The sequence for the target gene is obtained from a database.
- Nucleotides are added in the correct order to synthesise the correct base sequence.
- Protecting groups are added throughout the synthesis to make sure the correct nucleotides are added and no side branches are produced.

1Biological Molecules

1.1Monomers & Polymers

1.1.1Monomers & Polymers

1.1.2Condensation & Hydrolysis Reactions

1.2Carbohydrates

1.2.1Structure of Carbohydrates

1.2.2Types of Polysaccharides

1.2.3End of Topic Test - Monomers, Polymers and Carbs

1.2.4Exam-Style Question - Carbohydrates

1.2.5A-A* (AO3/4) - Carbohydrates

1.3Lipids

1.3.1Triglycerides & Phospholipids

1.3.2Types of Fatty Acids

1.3.3Testing for Lipids

1.3.4Exam-Style Question - Fats

1.3.5A-A* (AO3/4) - Lipids

1.4Proteins

1.4.1The Peptide Chain

1.4.2Investigating Proteins

1.4.3Primary & Secondary Protein Structure

1.4.4Tertiary & Quaternary Protein Structure

1.4.5Enzymes

1.4.6Factors Affecting Enzyme Activity

1.4.7Enzyme-Controlled Reactions

1.4.8End of Topic Test - Lipids & Proteins

1.4.9A-A* (AO3/4) - Enzymes

1.4.10A-A* (AO3/4) - Proteins

1.5Nucleic Acids

1.5.1DNA & RNA

1.5.2Nucleotides

1.5.3Polynucleotides

1.5.4DNA Replication

1.5.5Exam-Style Question - Nucleic Acids

1.5.6A-A* (AO3/4) - Nucleic Acids

1.6ATP

1.6.1Structure of ATP

1.6.2Hydrolysis of ATP

1.6.3Resynthesis of ATP

1.6.4End of Topic Test - Nucleic Acids & ATP

1.7Water

1.7.1Importance of Water

1.7.2Structure of Water

1.7.3Properties of Water

1.7.4A-A* (AO3/4) - Water

1.8Inorganic Ions

1.8.1Inorganic Ions

1.8.2End of Topic Test - Water & Inorganic Ions

2Cells

2.1Cell Structure

2.1.1Introduction to Cells

2.1.2Eukaryotic Cells & Organelles

2.1.3Eukaryotic Cells & Organelles 2

2.1.4Prokaryotes

2.1.5A-A* (AO3/4) - Organelles

2.1.6Methods of Studying Cells

2.1.7Microscopes

2.1.8End of Topic Test - Cell Structure

2.1.9Exam-Style Question - Cells

2.1.10A-A* (AO3/4) - Cells

2.2Mitosis & Cancer

2.2.1Mitosis

2.2.2Stages of Mitosis

2.2.3Investigating Mitosis

2.2.4Cancer

2.2.5A-A* (AO3/4) - The Cell Cycle

2.3Transport Across Cell Membrane

2.3.1Cell Membrane Structure

2.3.2A-A* (AO3/4) - Membrane Structure

2.3.3Diffusion

2.3.4Osmosis

2.3.5Active Transport

2.3.6End of Topic Test - Mitosis, Cancer & Transport

2.3.7Exam-Style Question - Membranes

2.3.8A-A* (AO3/4) - Membranes & Transport

2.3.9A-A*- Mitosis, Cancer & Transport

2.4Cell Recognition & the Immune System

2.4.1Immune System

2.4.2Phagocytosis

2.4.3T Lymphocytes

2.4.4B Lymphocytes

2.4.5Antibodies

2.4.6Primary & Secondary Response

2.4.7Vaccines

2.4.8HIV

2.4.9Ethical Issues

2.4.10End of Topic Test - Immune System

2.4.11Exam-Style Question - Immune System

2.4.12A-A* (AO3/4) - Immune System

3Substance Exchange

3.1Surface Area to Volume Ratio

3.1.1Size & Surface Area

3.1.2A-A* (AO3/4) - Cell Size

3.2Gas Exchange

3.2.1Single-Celled Organisms

3.2.2Multicellular Organisms

3.2.3Control of Water Loss

3.2.4Human Gas Exchange

3.2.5Ventilation

3.2.6Dissection

3.2.7Measuring Gas Exchange

3.2.8Lung Disease

3.2.9Lung Disease Data

3.2.10End of Topic Test - Gas Exchange

3.2.11A-A* (AO3/4) - Gas Exchange

3.3Digestion & Absorption

3.3.1Overview of Digestion

3.3.2Digestion in Mammals

3.3.3Absorption

3.3.4End of Topic Test - Substance Exchange & Digestion

3.3.5A-A* (AO3/4) - Substance Ex & Digestion

3.4Mass Transport

3.4.1Haemoglobin

3.4.2Oxygen Transport

3.4.3The Circulatory System

3.4.4The Heart

3.4.5Blood Vessels

3.4.6Cardiovascular Disease

3.4.7Heart Dissection

3.4.8Xylem

3.4.9Phloem

3.4.10Investigating Plant Transport

3.4.11End of Topic Test - Mass Transport

3.4.12A-A* (AO3/4) - Mass Transport

4Genetic Information & Variation

4.1DNA, Genes & Chromosomes

4.1.1DNA

4.1.2Genes

4.1.3Non-Coding Genes

4.1.4The Genetic Code

4.1.5A-A* (AO3/4) - DNA

4.2DNA & Protein Synthesis

4.2.1Protein Synthesis

4.2.2Transcription & Translation

4.2.3End of Topic Test - DNA, Genes & Protein Synthesis

4.2.4Exam-Style Question - Protein Synthesis

4.2.5A-A* (AO3/4) - Coronavirus Translation

4.2.6A-A* (AO3/4) - Transcription

4.2.7A-A* (AO3/4) - Translation

4.3Mutations & Meiosis

4.3.1Mutations

4.3.2Meiosis

4.3.3A-A* (AO3/4) - Meiosis

4.3.4Meiosis vs Mitosis

4.3.5End of Topic Test - Mutations, Meiosis

4.3.6A-A* (AO3/4) - DNA,Genes, CellDiv & ProtSynth

4.4Genetic Diversity & Adaptation

4.4.1Genetic Diversity

4.4.2Natural Selection

4.4.3A-A* (AO3/4) - Natural Selection

4.4.4Adaptations

4.4.5Investigating Natural Selection

4.4.6End of Topic Test - Genetic Diversity & Adaptation

4.4.7A-A* (AO3/4) - Genetic Diversity & Adaptation

4.5Species & Taxonomy

4.5.1Courtship Behaviour

4.5.2Phylogeny

4.5.3Classification

4.5.4DNA Technology

4.5.5A-A* (AO3/4) - Species & Taxonomy

4.6Biodiversity Within a Community

4.6.1Biodiversity

4.6.2Index of diversity

4.6.3Agriculture

4.6.4End of Topic Test - Species,Taxonomy& Biodiversity

4.6.5A-A* (AO3/4) - Species,Taxon&Biodiversity

4.7Investigating Diversity

4.7.1Genetic Diversity

4.7.2Quantitative Investigation

5Energy Transfers (A2 only)

5.1Photosynthesis

5.1.1Overview of Photosynthesis

5.1.2Photoionisation of Chlorophyll

5.1.3Production of ATP & Reduced NADP

5.1.4Cyclic Photophosphorylation

5.1.5Light-Independent Reaction

5.1.6A-A* (AO3/4) - Photosynthesis Reactions

5.1.7Limiting Factors

5.1.8Photosynthesis Experiments

5.1.9End of Topic Test - Photosynthesis

5.1.10A-A* (AO3/4) - Photosynthesis

5.2Respiration

5.2.1Overview of Respiration

5.2.2Anaerobic Respiration

5.2.3A-A* (AO3/4) - Anaerobic Respiration

5.2.4The Link Reaction

5.2.5The Krebs Cycle

5.2.6Oxidative Phosphorylation

5.2.7Respiration Experiments

5.2.8End of Topic Test - Respiration

5.2.9A-A* (AO3/4) - Respiration

5.3Energy & Ecosystems

5.3.1Biomass

5.3.2Production & Productivity

5.3.3Agricultural Practices

5.4Nutrient Cycles

5.4.1Nitrogen Cycle

5.4.2Phosphorous Cycle

5.4.3Fertilisers & Eutrophication

5.4.4End of Topic Test - Nutrient Cycles

5.4.5A-A* (AO3/4) - Energy,Ecosystems&NutrientCycles

6Responding to Change (A2 only)

6.1Nervous Communication

6.1.1Survival

6.1.2Plant Responses

6.1.3Animal Responses

6.1.4Reflexes

6.1.5End of Topic Test - Reflexes, Responses & Survival

6.1.6Receptors

6.1.7The Human Retina

6.1.8Control of Heart Rate

6.1.9End of Topic Test - Receptors, Retina & Heart Rate

6.2Nervous Coordination

6.2.1Neurones

6.2.2Action Potentials

6.2.3Speed of Transmission

6.2.4End of Topic Test - Neurones & Action Potentials

6.2.5Synapses

6.2.6Types of Synapse

6.2.7Medical Application

6.2.8End of Topic Test - Synapses

6.2.9A-A* (AO3/4) - Nervous Comm&Coord

6.3Muscle Contraction

6.3.1Skeletal Muscle

6.3.2Sliding Filament Theory

6.3.3Contraction

6.3.4Slow & Fast Twitch Fibres

6.3.5End of Topic Test - Muscles

6.3.6A-A* (AO3/4) - Muscle Contraction

6.4Homeostasis

6.4.1Overview of Homeostasis

6.4.2Blood Glucose Concentration

6.4.3Controlling Blood Glucose Concentration

6.4.4End of Topic Test - Blood Glucose

6.4.5Primary & Secondary Messengers

6.4.6Diabetes Mellitus

6.4.7Measuring Glucose Concentration

6.4.8Osmoregulation

6.4.9Controlling Blood Water Potential

6.4.10ADH

6.4.11End of Topic Test - Diabetes & Osmoregulation

6.4.12A-A* (AO3/4) - Homeostasis

7Genetics & Ecosystems (A2 only)

7.1Genetics

7.1.1Key Terms in Genetics

7.1.2Inheritance

7.1.3Linkage

7.1.4Multiple Alleles & Epistasis

7.1.5Chi-Squared Test

7.1.6End of Topic Test - Genetics

7.1.7A-A* (AO3/4) - Genetics

7.2Populations

7.2.1Populations

7.2.2Hardy-Weinberg Principle

7.3Evolution

7.3.1Variation

7.3.2Natural Selection & Evolution

7.3.3End of Topic Test - Populations & Evolution

7.3.4Types of Selection

7.3.5Types of Selection Summary

7.3.6Overview of Speciation

7.3.7Causes of Speciation

7.3.8Diversity

7.3.9End of Topic Test - Selection & Speciation

7.3.10A-A* (AO3/4) - Populations & Evolution

7.4Populations in Ecosystems

7.4.1Overview of Ecosystems

7.4.2Niche

7.4.3Population Size

7.4.4Investigating Population Size

7.4.5End of Topic Test - Ecosystems & Population Size

7.4.6Succession

7.4.7Conservation

7.4.8End of Topic Test - Succession & Conservation

7.4.9A-A* (AO3/4) - Ecosystems

8The Control of Gene Expression (A2 only)

8.1Mutation

8.1.1Mutations

8.1.2Effects of Mutations

8.1.3Causes of Mutations

8.2Gene Expression

8.2.1Stem Cells

8.2.2Stem Cells in Disease

8.2.3End of Topic Test - Mutation & Gene Epression

8.2.4A-A* (AO3/4) - Mutation & Stem Cells

8.2.5Regulating Transcription

8.2.6Epigenetics

8.2.7Epigenetics & Disease

8.2.8Regulating Translation

8.2.9Experimental Data

8.2.10End of Topic Test - Transcription & Translation

8.2.11Tumours

8.2.12Correlations & Causes

8.2.13Prevention & Treatment

8.2.14End of Topic Test - Cancer

8.2.15A-A* (AO3/4) - Gene Expression & Cancer

8.3Genome Projects

8.3.1Using Genome Projects

8.4Gene Technology

8.4.1Recombinant DNA

8.4.2Producing Fragments

8.4.3Amplification

8.4.4End of Topic Test - Genome Project & Amplification

8.4.5Using Recombinant DNA

8.4.6Medical Diagnosis

8.4.7Genetic Fingerprinting

8.4.8End of Topic Test - Gene Technologies

8.4.9A-A* (AO3/4) - Gene Technology

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