Difference between revisions of "Mukund's Brief Overview on Synthetic Biology Basics"

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== '''Some basics of DNA Cloning:''' ==
 
== '''Some basics of DNA Cloning:''' ==
  
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'''Recombinant DNA:'''
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Recombinant DNA is the general name for taking a piece of one DNA, and
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and combining it with another strand of DNA.
  
 
'''Plasmid:'''
 
'''Plasmid:'''
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When certain restriction enzymes cut DNA, they don't cut through both strands of DNA at the same point.The end that is left is called a sticky end. One strand is left with unpaired bases, which can later pair up with another sticky end created by the same enzyme.
 
When certain restriction enzymes cut DNA, they don't cut through both strands of DNA at the same point.The end that is left is called a sticky end. One strand is left with unpaired bases, which can later pair up with another sticky end created by the same enzyme.
  
 +
'''DNA Ligase:'''
 +
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A Secial type of ligase (EC 6.5.1.1) that can link together two DNA strands that have double-strand break (a break in both complementary strands of DNA)
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An overview of Transformation:
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Transformation is one of the three methods by which DNA cloning takes place today.
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 +
Method:
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1)A a piece of DNA to be inserted  into a vector is selected.
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 +
2)The DNA piece is cut with a restriction enzyme
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 +
3) The DNA to be inserted is ligated into the vector with DNA Ligase.
 +
enzyme and then ligate the DNA insert into the vector with DNA Ligase. The insert contains a selectable
 +
marker which allows for identification of recombinant molecules.
 
[[File:Dna cloning.gif|centre]]
 
[[File:Dna cloning.gif|centre]]

Revision as of 05:58, 21 May 2009

Q1- What will you see if you add mud to water?

A- Sedimentation and floating particles of mud; Darkish-colored water

Q2-What will you see if DNA if added to water?

A-Clear water.

How does the phenomenon of 'seeing' the particles of mud and not the DNA take place? This is because the particles of mud scatter light that results in the eye being able to view them. DNA, however,allows light to pass right through it, due to which it becomes invisible to the eye.

The discussion with Mukund
Scattering of light (sketch)
Flourescence of light (sketch)


There are three main ways by which light passes through an object:

a) Scattering

b) Absorption

c) Flouresence

  • There is one major problem with DNA, that is, its too small to see - even with high levels of of magnification.Some of the ways of attempting to see them are through:

a) Addition of dyes

b) Increasing the quantity of the sample observed



Some basics of DNA Cloning:

Recombinant DNA:

Recombinant DNA is the general name for taking a piece of one DNA, and and combining it with another strand of DNA.

Plasmid:

A DNA molecule that is separate from the chromosomal DNA and is capable of replicating independently.This is a small, circular piece of DNA, usually a few thousand base pairs long, which can be found in bacteria

Palindrome:

A sequence that reads the same in the forward and backward direction.

The Sequence: ATGCAT

Itc complement :TACGTA

It is due to this characteristic of the DNA sequences that they can bind to other sequences.

Restriction enzymes:

Enzymes responsible for cuting DNA strands.They recognize a specific, short sequence of DNA and cut the DNA at that point. Different restriction enzymes recognize and cut different sequences.

Sticky end:

When certain restriction enzymes cut DNA, they don't cut through both strands of DNA at the same point.The end that is left is called a sticky end. One strand is left with unpaired bases, which can later pair up with another sticky end created by the same enzyme.

DNA Ligase:

A Secial type of ligase (EC 6.5.1.1) that can link together two DNA strands that have double-strand break (a break in both complementary strands of DNA) An overview of Transformation:

Transformation is one of the three methods by which DNA cloning takes place today.

Method: 1)A a piece of DNA to be inserted into a vector is selected.

2)The DNA piece is cut with a restriction enzyme

3) The DNA to be inserted is ligated into the vector with DNA Ligase. enzyme and then ligate the DNA insert into the vector with DNA Ligase. The insert contains a selectable marker which allows for identification of recombinant molecules.

Dna cloning.gif