This document discusses cryptic satellite DNA. It begins by introducing satellite DNA as highly repetitive sequences that form distinct bands during density gradient centrifugation. Cryptic satellite DNA does not form distinct bands due to base methylation. The document then provides two examples of cryptic satellite DNA: 1) In Drosophila virilis, there are three major satellites and a cryptic one. 2) In a hermit crab, one major very highly repeated DNA and three minor variants comprising inverted repeats account for 30% of its genome. These minor variants are considered cryptic satellites.
Genomic library and shotgun sequencing. It includes the topics about genomic library,construction method, its uses and applications, shotgun sequencing, difference between random and whole genome sequencing, its advantages and disadvantages etc.
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A DNA library is a collection of cloned restriction fragments of the DNA of an organism.
Two kinds of libraries will be discussed: genomic libraries and complementary DNA (cDNA) libraries.
Genomic libraries ideally contain a copy of every DNA nucleotide sequence in the genome.
In contrast, cDNA libraries contain those DNA sequences that appear as mRNA molecules, and these differ from one cell type to another.
Genomic library and shotgun sequencing. It includes the topics about genomic library,construction method, its uses and applications, shotgun sequencing, difference between random and whole genome sequencing, its advantages and disadvantages etc.
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A DNA library is a collection of cloned restriction fragments of the DNA of an organism.
Two kinds of libraries will be discussed: genomic libraries and complementary DNA (cDNA) libraries.
Genomic libraries ideally contain a copy of every DNA nucleotide sequence in the genome.
In contrast, cDNA libraries contain those DNA sequences that appear as mRNA molecules, and these differ from one cell type to another.
As a periodontist, it is of utmost importance to understand the genetic basis of inheritance in periodontal diseases be able to relate to the various polymorphisms associated with periodontal diseases. This ppt presents the basics of genetics from the point of view of future understanding of polymorphisms related to periodontal diseases.
DNA replication is the most important process central dogma in the molecular genetics. So i hope this power point presentation useful to the students of B.Sc Agriculture and M.Sc Genetics and Plant Breeding.
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Gregorius and Namkoong (1986) defined Separability as the property which is that cultivar effect is separable from environmental effect so that there is no rank.
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2. INTRODUCTION
DNA is a polymers of polynucleotide sequences.
Its found in living organism, either may be double helical or may be circular form.
Double helical DNA Circular DNA
3. DNA subdivided on the basis of sequences of nucleotide in two type :-
1.Non repetitive DNA (unique DNA) :- single copy per genome in living organism.
2. repetitive DNA :- few to million copy per genome.
Also subdivided on the basis of no. of copies of nucleotide sequences :-
A. Highly repetitive DNA :- million copies per genome.
B. Moderately repetitive DNA :- few to thousand copies per genome.
Information on repetitive DNA has been gathered through Density Gradient
Centrifugation Method.
4.
5. SATELLITE DNA
Some very short highly repetitive sequences (repeated many time in tandem) differ in
density from the remaining chromosomal DNA and are separable as minor band in
CsCl2 Density Gradient Centrifugation, Called as SATELLITE DNA.
An important feature of satellite is that they represent very long stretches of DNA of very
low sequence complexity, within constancy of sequences can be maintained.
6. Where are the blocks of highly repetitive DNA located in the genome?
It is located in heterochromatin regions of chromosomes, Heterochromatin is commonly
found at centromeres.
Why?
(the regions where the kinetochores are formed at mitosis and meiosis for controlling
chromosome movement).
The centromeric location of satellite DNA suggests that it has some structural function in
the chromosome.
This function could be connected with the process of chromosome segregation.
7.
8.
9.
10. CRYPTIC SATELLITE DNA
Generally single satellite sequences found , but some time multiple satellite sequences
can be there, which may anomalous behavior in CsCl2 Density Gradient Centrifugation
due to base methylation so that cannot be separated from the main-band DNA so it is
called CRYPTIC SATELLITE DNA.
cryptic and apparent satellites usually account for all the large tandemly repeated blocks
of highly repetitive DNA. When a genome has more than one type of highly repetitive
DNA, each exists in its own satellite block although sometimes different blocks are
adjacent.
11.
12. Minisatellite:-
Some sequences that resemble satellite DNA consists of tandem repeats of a short
sequences of 2 to 8 bp, the repeats being identical.
Variation in this type of DNA arises due mainly to differences in the number of repeat
units, which is highly variable.
These DNAs are called minisatellite or VNTRs (variable number of tandem repeats)
and are used for fingerprinting and genome mapping.
13. Examples:-
1.Drosophila virilis,
three major satellites, viz., ACAAACT, ATAAACT, ACAAATT, and a cryptic satellite DNA
(AATATAG) make up 40% of the genome.
but satellites in other species may have unrelated sequences.
Each satellite has arisen by a lateral amplification of a very short sequences, which may
be a variant of a preexisting satellite or may have Some other origin.
14.
15. 2.Mus musculus
Satellite DNA consists mainly of a repeating unit of 234 bp.
This fragment is composed of two smaller sub units of 117 bp each.
Each 117 bp subunit contains two smaller sequences of 58 and 59 (one T added to the
second subunit) bp.
Each 58/59 bp subunit, in turn, has two subunits called α & β; the α sequences have an
insertion of one C ,while β sequences have an insertion of a 3 bp sequences.
The analysis of 58/59 bp reveals consensus of 9 bases, which is the basic repeating
unit.
This repeating sequences is supposed to have become amplified by a process called
saltatory replication to yield 54 bp subunit, which became 58/59 bp by addition. The α
& β subunits of the 58/59 bp subunit diverged due to the accumulation of mutation.
Subsequently, the 58/59 bp subunit underwent saltatory replication as a unit to yield
the much larger 234 bp repeating unit.
Mutation accumulation would have resulted in further divergence among the repeating
units.
16.
17. Cryptic Satellites in Inverted Repeats Comprise 30% of
the Genome of a HERMIT CRAB:-
One major very highly repeated (VHR) DNA (repeat unit = 156 bp), a family of three
minor VHR DNAs (repeat units = 71-74 bp), and a number of trace components account
for almost 30% of the genome of a hermit crab.
The repeat units of the three minor variants are defined by identical 14-bp G + C rich
inverted repeats that might form cruciform.
Two copies of the repeat unit (CCTA) of one of two patent satellites of this crab occur at
the center of one in seven of the G + C- rich inverted repeats;
copies of the other patent satellite are found in main component DNA.
18. The sequences of both the major and minor VHR DNAs are characterized by short
tracts of An and/or Tn(n = 4-7) residues whose presence would permit the formation of
perfectly matched stems separated by loops of 8-16 bp.
The An and/or Tn tracts are interspersed with segments of G + C-rich DNA and are
arranged differently in the major and minor VHR DNAs.
Although the repeat units of the major and the three minor VHR DNAs are arranged in
tandem, the composition and sequence of their bases are such that they do not form
distinct bands in CsCl2 gradients. They are called Cryptic Satellites.
Other example of cryptic satellite DNA :-
Complex organization of a cryptic satellite DNA in the
genome of the marine invertebrate Rapana
thomasiana (Gastropoda).