Successfully reported this slideshow.
We use your LinkedIn profile and activity data to personalize ads and to show you more relevant ads. You can change your ad preferences anytime.
1
Presented by
Dr. Akshay S. Joshi
Advanced Techniques
In Molecular
Cytogenetics
Karyotyping, FISH, CGH, SKY
Introduction
History
Importance of molecular cytogenetical
techniques
Molecular cytogenetical techniques
and their applica...
What Is Cytogenetics……?
Genetics - The branch of science which deals with
the study of heredity and the variation is calle...
Human have
tendency to
reveal the
secrets behind
every thing…..
Molecular
Biology
Molecular Cytogenetics
Molecular Cytogenetics
Cytogenetics
Cytology
Genetics
Molecular Cytogenetics
It i...
• The first illustration of human chromosomes
• First used the term mitosis1882
• Waldeyer introduced the term Chromosome
...
• Langer develop for the non isotope labelling
of DNA by conjugation of biotin1981
• Albertson detected biotin labelled pr...
Why Molecular Cytogenetical
Techniques
• Molecular cytogenetical techniques are use to
detect chromosomal aberrations.
• T...
Causes Of Chromosomal
Abnormalities
Chromosomes Number
Chromosomes Size
The Arrangement Of
The Chromosome
Segment
No. Of C...
Variation In Chromosome Number
• A general term for the variation in the no.
• Aneuploidy
• Chromosomes no. that are not e...
• Euploidy / Polyploidy
• Increase in chromosomes numbers due to 1 or more
complete sets of Chromosomes is called as euplo...
Variation In No. Of Chromosome
Segment
• Deletion
• It is a common phenomenon of a chromosome to break.
Two types of delet...
Variation in chromosome
morphology
1. Isochromosomes
2. Bridge-breakage –fusion-bridge cycles:
3. Ring chromosome:
ABNORMALITIES OF GENES/DNA
1. Point mutation
2. Gross mutation
3. Nonsense mutation
4. Silent mutation
5. Missense mutatio...
Genetically abnormal animals
Partial Hypotrichosis Complete Hypotrichosis
congenital alopecia and anaemia presance of mark...
Effect of Genetic Diseases on Livestock : an
Indian Scenario
AI
Chromosomally
abnormal bull
Use of chromosomally
abnormal ...
MATING WITHOUT TESTING
NS
WITH
CARRIER
BULL
AFTER
GENETIC
TESTING
AFETR GENETIC TESTINGAFETR GENETIC TESTING
GENETICALLY C...
Worldwide reports about the economic losses
due to molecular genetic disorders
• BLAD (Bovine Leukocyte Adhesion Deficienc...
Genetic disorders reported in India
• 1;29 CFT in Jersey cross cow in Chennai (Thiagarajan et al., 1990)
• 1;29 CFT in Jer...
PREVENTION IS BETTER THAN
CURE….
!!
Prevention and Control
• Inherited genetic disorders affect all kinds of farm animals.
• The physiological defects arising...
Which Are The Molecular Cytogenetic
Techniques…..?
Karyotyping
Banding
Techniques
Fluroscent In Situ
Hybridization
Compara...
What Is Karyotyping
• The study of whole sets of chromosomes is sometimes
known as karyology.
• The chromosomes are depict...
What karyotyping does…..?
• These techniques produce a characteristic pattern of
contrasting dark and light transverse ban...
How To Do Karyotyping……?
Arrange them in order.
Matched by banding patterns
Cut out
Enlarged
Picture taken
Chromosomes sta...
Which are chromosomal banding
techniques……?
G – banding
R - banding
Q – banding
T – banding
Silver staining
G-banding
• G banding is obtained
with Giemsa stain following
digestion of chromosomes
with trypsin.
• It yields a series ...
R-banding
• is the reverse of G-banding
(the R stands for "reverse").
• The dark regions are
euchromatic (guanine-
cytosin...
Arrangement of chromosomes
• Picture shows abrupt
arrangement haploid
chromosome.
• Then scientist with
experience cut out...
Applications Of Karyotyping
• Karyotype show the chromosomal makeup of an
individual.
• Knowing the number of chromosomes ...
Limitations Of Karyotyping
• Expert technician is require to
arrange the chromosome.
• Structural abnormalities can not be...
Fluroscent in situ hybridization
(FISH)
What is FISH…..?
• Fluorescence in situ hybridization (FISH)
is a cytogenetic technique that
uses fluorescent probes that ...
What are probes
• probe is a synthesized fragment of DNA or RNA of
variable length which can be radioactively labeled
• Th...
Fluroscent In Situ Hybridization
• Step I – Denaturation
Conversion of double stranded dna in to single stranded
dna
• St...
Slide With Metaphase
Chromosomes
Denaturation Of DNA
Addition Of Probe
Hybridization
Washing And
Addition Of Counter
Stain...
Types of FISH….?
Interphase FISH
Telomeric FISH
RNA in situ
hybridization
Prime in situ
labelling
COBRA FISH
SKY FISH
M- F...
Application
• FISH generally used compliment classical staining technique
• Substitute for chromosome identification at me...
• Helps in detection of single gene disorder and
presence or absence of particular gene on
chromosome.
• By using probe ch...
Comparative Genomic Hybridization
(CGH)
• Comparative genomic hybridization is a molecular
cytogenetic method for analyzin...
Software analysis
Fluroscence detector image analysis
Hybridization
Selection and application of probe
Isolation of DNA fr...
Standard Patient Abnormal
(Probe)
Fluorescent Detector
(Probe) (Probe)
Duplication Deletion
Standard Abnormal Standard Abnormalnormal normal
Computer analysis
Software analysis
Graphical image by softwareFluorescence detector set
Fluorescence detector
Application of CGH
• This technology was first developed as a research tool for the investigation of
genomic alterations i...
Spectral Karyotyping (SKY)
What is SKY…?
• Spectral karyotyping is cytogenetical technique used
to simultaneously visualize all the pairs of
chromoso...
How to do SKY….?
Visualization
Image Acquisition
Washes And Detection
Application Of Dye
Hybridization
Probe Denaturation
...
Spectral Karyotype Observation
Some Abnormalities Seen In SKY
Translocation
Application of SKY
• This technology allows the isolation of structural
chromosome abnormalities
• which then allows deter...
Conclusion
• Chromosomal genetic abnormalities are the hidden
cause for the huge economic losses.
• Molecular cytogenetic ...
References
Ashutosh H., M. Jain, I. Chaudhary, N. Gupta and M. Kabra 2013 Fluorescence in situ hybridization (FISH) using ...
Advanced Techniques In Molecular Cytogenetics Karyotyping, FISH, CGH, SKY
Advanced Techniques In Molecular Cytogenetics Karyotyping, FISH, CGH, SKY
Advanced Techniques In Molecular Cytogenetics Karyotyping, FISH, CGH, SKY
Upcoming SlideShare
Loading in …5
×

Advanced Techniques In Molecular Cytogenetics Karyotyping, FISH, CGH, SKY

12,276 views

Published on

Advanced Techniques In Molecular Cytogenetics
Karyotyping, FISH, CGH, SKY

Published in: Science
  • Hi there! I just wanted to share a list of sites that helped me a lot during my studies: .................................................................................................................................... www.EssayWrite.best - Write an essay .................................................................................................................................... www.LitReview.xyz - Summary of books .................................................................................................................................... www.Coursework.best - Online coursework .................................................................................................................................... www.Dissertations.me - proquest dissertations .................................................................................................................................... www.ReMovie.club - Movies reviews .................................................................................................................................... www.WebSlides.vip - Best powerpoint presentations .................................................................................................................................... www.WritePaper.info - Write a research paper .................................................................................................................................... www.EddyHelp.com - Homework help online .................................................................................................................................... www.MyResumeHelp.net - Professional resume writing service .................................................................................................................................. www.HelpWriting.net - Help with writing any papers ......................................................................................................................................... Save so as not to lose
       Reply 
    Are you sure you want to  Yes  No
    Your message goes here
  • Don't forget another good way of simplifying your writing is using external resources (such as ⇒ www.HelpWriting.net ⇐ ). This will definitely make your life more easier
       Reply 
    Are you sure you want to  Yes  No
    Your message goes here
  • Copas Url to Download PDF eBook === http://capitolcurrents.com/0749675527-fish.html
       Reply 
    Are you sure you want to  Yes  No
    Your message goes here
  • How To Pick Out The Best Battery For a Solar Panel System, Battery Bank, or Off-Grid System ♣♣♣ http://t.cn/AiFAGBwT
       Reply 
    Are you sure you want to  Yes  No
    Your message goes here
  • DOWNLOAD THIS BOOKS INTO AVAILABLE FORMAT (2019 Update) ......................................................................................................................... ......................................................................................................................... Download Full PDF EBOOK here { https://soo.gd/irt2 } ......................................................................................................................... Download Full EPUB Ebook here { https://soo.gd/irt2 } ......................................................................................................................... Download Full doc Ebook here { https://soo.gd/irt2 } ......................................................................................................................... Download PDF EBOOK here { https://soo.gd/irt2 } ......................................................................................................................... Download EPUB Ebook here { https://soo.gd/irt2 } ......................................................................................................................... Download doc Ebook here { https://soo.gd/irt2 } ......................................................................................................................... ......................................................................................................................... ................................................................................................................................... eBook is an electronic version of a traditional print book THIS can be read by using a personal computer or by using an eBook reader. (An eBook reader can be a software application for use on a computer such as Microsoft's free Reader application, or a book-sized computer THIS is used solely as a reading device such as Nuvomedia's Rocket eBook.) Users can purchase an eBook on diskette or CD, but the most popular method of getting an eBook is to purchase a downloadable file of the eBook (or other reading material) from a Web site (such as Barnes and Noble) to be read from the user's computer or reading device. Generally, an eBook can be downloaded in five minutes or less ......................................................................................................................... .............. Browse by Genre Available eBooks .............................................................................................................................. Art, Biography, Business, Chick Lit, Children's, Christian, Classics, Comics, Contemporary, Cookbooks, Manga, Memoir, Music, Mystery, Non Fiction, Paranormal, Philosophy, Poetry, Psychology, Religion, Romance, Science, Science Fiction, Self Help, Suspense, Spirituality, Sports, Thriller, Travel, Young Adult, Crime, Ebooks, Fantasy, Fiction, Graphic Novels, Historical Fiction, History, Horror, Humor And Comedy, ......................................................................................................................... ......................................................................................................................... .....BEST SELLER FOR EBOOK RECOMMEND............................................................. ......................................................................................................................... Blowout: Corrupted Democracy, Rogue State Russia, and the Richest, Most Destructive Industry on Earth,-- The Ride of a Lifetime: Lessons Learned from 15 Years as CEO of the Walt Disney Company,-- Call Sign Chaos: Learning to Lead,-- StrengthsFinder 2.0,-- Stillness Is the Key,-- She Said: Breaking the Sexual Harassment Story THIS Helped Ignite a Movement,-- Atomic Habits: An Easy & Proven Way to Build Good Habits & Break Bad Ones,-- Everything Is Figureoutable,-- What It Takes: Lessons in the Pursuit of Excellence,-- Rich Dad Poor Dad: What the Rich Teach Their Kids About Money THIS the Poor and Middle Class Do Not!,-- The Total Money Makeover: Classic Edition: A Proven Plan for Financial Fitness,-- Shut Up and Listen!: Hard Business Truths THIS Will Help You Succeed, ......................................................................................................................... .........................................................................................................................
       Reply 
    Are you sure you want to  Yes  No
    Your message goes here

Advanced Techniques In Molecular Cytogenetics Karyotyping, FISH, CGH, SKY

  1. 1. 1 Presented by Dr. Akshay S. Joshi Advanced Techniques In Molecular Cytogenetics Karyotyping, FISH, CGH, SKY
  2. 2. Introduction History Importance of molecular cytogenetical techniques Molecular cytogenetical techniques and their applications Conclusion
  3. 3. What Is Cytogenetics……? Genetics - The branch of science which deals with the study of heredity and the variation is called as genetics. Cytology - The branch of science which deals with the study ofstructure and function of cell and cell organelles is celled as cytology. Cytogenetic - The branch of genetics which deals with the study of cell and structure and function of the chromosomes is called as cytogenetic.
  4. 4. Human have tendency to reveal the secrets behind every thing…..
  5. 5. Molecular Biology Molecular Cytogenetics Molecular Cytogenetics Cytogenetics Cytology Genetics Molecular Cytogenetics It involves the combination of molecular biology and cytogenetic. In general this involves the use of a series of techniques in which DNA probes are labeled with different colored fluorescent tags to visualize one or more specific regions of the genome.
  6. 6. • The first illustration of human chromosomes • First used the term mitosis1882 • Waldeyer introduced the term Chromosome • Greek words for colored body 1888 • Sutton combined two disciplines, cytology and genetics as Cytogenetics1902 • Levitsky termed The ordered arrangement of chromosomes as karyotype1924 • Caspersson and co-workers reported a unique banding pattern • Drets and Shaw produces banding pattern by Giemsa stain 1971 • Pardue and Gall demonstrated in situ hybridization. 1969 • Tjio and Levan start human genome project1956
  7. 7. • Langer develop for the non isotope labelling of DNA by conjugation of biotin1981 • Albertson detected biotin labelled probes1983 • Guan use chromosome microdissection for whole chromosome painting probes (WCPs)1994 • Speicher develops multiplex fluorescence in situ hybridization (M-FISH) • Schröck develops a spectral karyotyping (SKY) 1996 • Tanke combined Binary RAtio labeling-FISH (COBRA FISH) • Henegariu color-changing karyotyping (CCK) • Chudoba developed multicolor banding (MCB) 1999 • Aurich-costa IPM-FISH2001
  8. 8. Why Molecular Cytogenetical Techniques • Molecular cytogenetical techniques are use to detect chromosomal aberrations. • There are many chromosomal aberrations are observe in the population.
  9. 9. Causes Of Chromosomal Abnormalities Chromosomes Number Chromosomes Size The Arrangement Of The Chromosome Segment No. Of Chromosome Segment Variation in chromosome morphology Variation In
  10. 10. Variation In Chromosome Number • A general term for the variation in the no. • Aneuploidy • Chromosomes no. that are not exact multiples of n. • It is further classified as 1. Monosomic: 2. Trisomic: 3. Tetrasomic: 4. Double trisomic: 5. Nullosomic:
  11. 11. • Euploidy / Polyploidy • Increase in chromosomes numbers due to 1 or more complete sets of Chromosomes is called as euploidy. 1. Monoploid: 2. Diploid: 3. Triploid: 4. Tetraploid: 5. Autotetraploid: 6. Polyploid:
  12. 12. Variation In No. Of Chromosome Segment • Deletion • It is a common phenomenon of a chromosome to break. Two types of deletions are possible 1. Terminal 2. Intercalary • Duplication: • Large or small piece of chromosome containing extra blocks of genes due to duplication are found in many individuals or even in races.
  13. 13. Variation in chromosome morphology 1. Isochromosomes 2. Bridge-breakage –fusion-bridge cycles: 3. Ring chromosome:
  14. 14. ABNORMALITIES OF GENES/DNA 1. Point mutation 2. Gross mutation 3. Nonsense mutation 4. Silent mutation 5. Missense mutation 6. Neutral mutation 7. Frameshift mutation 8. Structural mutation
  15. 15. Genetically abnormal animals Partial Hypotrichosis Complete Hypotrichosis congenital alopecia and anaemia presance of marked wrinkling of the skin
  16. 16. Effect of Genetic Diseases on Livestock : an Indian Scenario AI Chromosomally abnormal bull Use of chromosomally abnormal semen Infertility, repeat breeding, genetic disorder Huge disaster Dead and live sperm, sperm motility, semen volume Solution Genetic testing of animal
  17. 17. MATING WITHOUT TESTING NS WITH CARRIER BULL AFTER GENETIC TESTING AFETR GENETIC TESTINGAFETR GENETIC TESTING GENETICALLY CLEAN POPULATION Consequences
  18. 18. Worldwide reports about the economic losses due to molecular genetic disorders • BLAD (Bovine Leukocyte Adhesion Deficiency). It was estimated that 16,000 calves were born with BLAD each year. Hence, the average economic loss per calf was roughly 300 US dollors. • Prevalence of BLAD in Indian HF crossbred cattle population is an alarming situation due to consistent reports from 1999 to 2013. • Factor XI deficiency (FXID) is responsible for repeat breeding in carriers leading to the financial losses . In Turkey (Akyuz et al., 2012) reported an extra $ 324 financial loss compared with a normally fertile cow due to this syndrome. • CVM ( Complex Vertebral Malformation ) carriers in India and worldwide much more higher than other four disorders i.e. BLAD, DUMPS, Citrullinemia and Factor XI • In 2012 in New Zealand Livestock Improvement Corporation (LIC) , a farmer cooperative that sells bull semen was been warned for supplying defective semen to farmers who produced 1500 defective calves with genetic defect leading to excessive hairiness and intolerance to heat ultimately reducing milk producing ability.
  19. 19. Genetic disorders reported in India • 1;29 CFT in Jersey cross cow in Chennai (Thiagarajan et al., 1990) • 1;29 CFT in Jersey cross bull in MS used through AI for 9 years (Chauhan et al.2009) • 7;16 CFT in HF cross bull under use through AI in KDC Mumbai ( Biotech lab report 1999-2000) • 7;16 CFT in HF cross bull before use through AI in MS (Patel,1999) • 1;9 CFTs in Gir bull in Gujarat State ( Paderi et al., 2011b) • 16;20 CFT in Deoni bull calf in 16;20 in Karanataka State ( Paderi et al., 2011a) • 8;29 CFT in one Gaolao Bull and it’s 12 progeny in MS ( Umrikar et al., 2013)
  20. 20. PREVENTION IS BETTER THAN CURE…. !!
  21. 21. Prevention and Control • Inherited genetic disorders affect all kinds of farm animals. • The physiological defects arising from inherited disorders have negative impact on health and productivity of farm animals and lead to economic loss in the dairy cattle industry. • Spread of undesirable genetic disorder is hastened by the artificial insemination programme. • Therefore, diagnosis or detection of chromosomal abnormalities is required to diminish the risk of dissemination of recessive disorders using advance molecular cytogenetical techniques in the dairy industry.  The mandatory provisions of advance molecualr cytogenetical techniques.  The semen of every bull in commercial farms should be screened out using molecular techniques.  A Genetic Investigation Laboratory has been established in Maharashtra Animal and Fishery Sciences University at Bombay Veterinary College, Mumbai with support from Government of India under Rashtriya Krishi Vikas Yojana in 2012.
  22. 22. Which Are The Molecular Cytogenetic Techniques…..? Karyotyping Banding Techniques Fluroscent In Situ Hybridization Comparative Genomic Hybridization Spectral Karyotyping Vartual Karyotyping
  23. 23. What Is Karyotyping • The study of whole sets of chromosomes is sometimes known as karyology. • The chromosomes are depicted (by rearranging a photomicrograph) in a standard format known as a karyogram or idiogram • Karyotyping is a test to examine chromosomes in a sample of cells. This test can help identify genetic problems as the cause of a disorder or disease
  24. 24. What karyotyping does…..? • These techniques produce a characteristic pattern of contrasting dark and light transverse bands on the chromosomes. • Banding makes it possible to identify homologous chromosomes by visualization of chromosomes. • Banding of homologous chromosomes allows chromosome segments and rearrangements to be identified. • it helps to detect the chromosomal abnormalities and chromosomal alterations. • The most widely used banding methods are G- banding (Giemsa-banding) and R-banding (reverse- banding).
  25. 25. How To Do Karyotyping……? Arrange them in order. Matched by banding patterns Cut out Enlarged Picture taken Chromosomes stained Cell fixed in metaphase
  26. 26. Which are chromosomal banding techniques……? G – banding R - banding Q – banding T – banding Silver staining
  27. 27. G-banding • G banding is obtained with Giemsa stain following digestion of chromosomes with trypsin. • It yields a series of lightly and darkly stained bands — The dark regions tend to be heterochromatic, late-replicating and AT rich. • The light regions tend to be euchromatic, early-replicating and GC rich. • This method will normally produce 300–400 bands on chromosome.
  28. 28. R-banding • is the reverse of G-banding (the R stands for "reverse"). • The dark regions are euchromatic (guanine- cytosine rich regions) • and the bright regions are heterochromatic (thymine- adenine rich regions).
  29. 29. Arrangement of chromosomes • Picture shows abrupt arrangement haploid chromosome. • Then scientist with experience cut out each chromosome and arrange them in order according to their size and pair.
  30. 30. Applications Of Karyotyping • Karyotype show the chromosomal makeup of an individual. • Knowing the number of chromosomes is essential for identifying chromosomal variations that cause genetic disorders. • Correct number of chromosomes and chromosomal abnormality in numbers can be detect. • Correct size and shape of chromosomes is visualize in this technique. • We can identify Gender of unknown sample by this technique.
  31. 31. Limitations Of Karyotyping • Expert technician is require to arrange the chromosome. • Structural abnormalities can not be detected by this technique. • By observing karyograph defects like deletion, duplication and many other can not be detected.
  32. 32. Fluroscent in situ hybridization (FISH)
  33. 33. What is FISH…..? • Fluorescence in situ hybridization (FISH) is a cytogenetic technique that uses fluorescent probes that bind to only those parts of the chromosome with a high degree of sequence complementarity • It is used to detect and localize the presence or absence of specific DNA sequences on chromosomes . • FISH is often used for finding specific features in DNA for use in genetic counseling, medicine, and species identification
  34. 34. What are probes • probe is a synthesized fragment of DNA or RNA of variable length which can be radioactively labeled • The size may be varies from 100-1000 bases long. • The probe thereby hybridizes to single-stranded nucleic acid (DNA or RNA) whose base sequence allows probe- target base pairing due to complementarity between the probe and target. • the probe is tagged (or "labeled") with a molecular marker of either radioactive or fluorescent molecules.
  35. 35. Fluroscent In Situ Hybridization • Step I – Denaturation Conversion of double stranded dna in to single stranded dna • Step II – Hybridization Application of probe DNA to slide & overnight incubation at 37°C Binding of probe DNA to trarget DNA. • Step III – Post hybridisation washing & detection Washing of unbound probe DNA. Application of counter stain & • Step IV – counter stain Application of counter stain. • Step V – Visualization visualization using fluorescence microscopy.
  36. 36. Slide With Metaphase Chromosomes Denaturation Of DNA Addition Of Probe Hybridization Washing And Addition Of Counter Stain Visualization
  37. 37. Types of FISH….? Interphase FISH Telomeric FISH RNA in situ hybridization Prime in situ labelling COBRA FISH SKY FISH M- FISH Fibre FISH
  38. 38. Application • FISH generally used compliment classical staining technique • Substitute for chromosome identification at metaphase or interphase • Useful in several clinical settings to determine prognosis and detection of genetic abnormalities like anupolidy, characteristic gene fusion, cross of chromosomal region or whole chromosom. • Discrete information is obtained from each cell
  39. 39. • Helps in detection of single gene disorder and presence or absence of particular gene on chromosome. • By using probe chromosomal material of unknown or uncertain organism can be identified • FISH is powerful technique use in detection of chromosomal abnormalities • Most significance advance in both research like gene mapping or identification of noval oncogenes or genetic abberation and diagnosis of haematological malignancies and solid tumor.
  40. 40. Comparative Genomic Hybridization (CGH) • Comparative genomic hybridization is a molecular cytogenetic method for analyzing copy number variations with the help of hybridization technique.
  41. 41. Software analysis Fluroscence detector image analysis Hybridization Selection and application of probe Isolation of DNA from Test Tissue and Reference Tissue Metaphase Slide Preparation Same as FISH CGH
  42. 42. Standard Patient Abnormal (Probe) Fluorescent Detector (Probe) (Probe)
  43. 43. Duplication Deletion Standard Abnormal Standard Abnormalnormal normal
  44. 44. Computer analysis Software analysis Graphical image by softwareFluorescence detector set Fluorescence detector
  45. 45. Application of CGH • This technology was first developed as a research tool for the investigation of genomic alterations in cancer. • It allows for a high-resolution evaluation of DNA copy number alterations associated with chromosome abnormalities. • It provides clinicians with a powerful tool to use in their increasingly sophisticated diagnostic capabilities. • The use of CGH is considered EXPERIMENTAL AND/OR INVESTIGATIONAL when performed in the absence of symptoms or high risk factors for a genetic disease or when knowledge of genetic status will not affect treatment decisions or screening for the disease. • Screening for prenatal gene mutations in fetuses without structural abnormalities or testing products of conception after AI. • Diagnosis of melanoma. • It helps in detection of balanced rearrangements of chromosomes and for comparison of normal and suspected DNA samples.
  46. 46. Spectral Karyotyping (SKY)
  47. 47. What is SKY…? • Spectral karyotyping is cytogenetical technique used to simultaneously visualize all the pairs of chromosome in an organism in different colors. • The sky technique is useful for identifying chromosomal abnormalities. • We can arrange the chromosomes according to their number just by visualization of different colours aquire by the chromosomes.
  48. 48. How to do SKY….? Visualization Image Acquisition Washes And Detection Application Of Dye Hybridization Probe Denaturation Denaturation Of DNA Chromosome Preparation From Tissue Same as FISH
  49. 49. Spectral Karyotype Observation
  50. 50. Some Abnormalities Seen In SKY Translocation
  51. 51. Application of SKY • This technology allows the isolation of structural chromosome abnormalities • which then allows determination of the precise molecular address of chromosome breakpoints associated with deletions translocations and insertion. • SKY can discern the aberrations that can’t be detected very well by conventional banding technique and Fluorescent in situ hybridization (FISH). • It allows visualization of all chromosomes in different colours on same platform which is very easy to detect chromosomal abnormalities.
  52. 52. Conclusion • Chromosomal genetic abnormalities are the hidden cause for the huge economic losses. • Molecular cytogenetic techniques like FISH, CGH and SKY are the available advanced diagnostic tools to detect such chromosomal abnormalities and to prevent the spread in the population which indirectly avoid the economic consequences.
  53. 53. References Ashutosh H., M. Jain, I. Chaudhary, N. Gupta and M. Kabra 2013 Fluorescence in situ hybridization (FISH) using non-commercial probes in the diagnosis of clinically suspected microdeletion syndromes. Indian J Med Res 138, pp 135-142. Christine M. (2008) Spectral Karyotyping (Sky) Establishes Chromosomal Homologies Between A New World Monkey (Pithecia Pithecia) And Humans (Homo Sapiens). TCNJ Journal Volume X : 1 – 12. Emanuela V. V. and J. M. Bridger (2008) FISH glossary: an overview of the fluorescence in situ hybridization technique. BioTechniques Vol. 45 (4): 385 – 405. George I. and O. Arisaka (2012) Chromosome Analysis Using Spectral Karyotyping (SKY). Cell Biochem Biophys 62:13–17. Jeffrey M. and R. H. Singer (2003) Fluorescence in situ hybridization: past, present and future. Journal of Cell Science 116 (14) 2833 – 2838. Kakazu N. and T. Abe (2006) Multicolor banding technique, spectral color banding (SCAN): new development and applications. Cytogenet Genome Res 114: 250–256. Malcolm A. F. S. (2015) History and evolution of cytogenetics. Molecular Cytogenetics 8 (19): 2– 8. R. Raff and G. Schwanitz (2001) Fluorescence in situ Hybridization General Principles and Clinical Application with Special Emphasis to Interphase Diagnostics. IJHG 1(1): 65-75. Regine S., E. Schrock, T. Ried and G. Schwanitz (2004) Spectral Karyotyping (SKY) permits the Characterisation of a de Novo unbalanced Translocation 46, XY, der(14)t(12;14). Int J Hum Genet, 4 (3): 173-178. Wendy A. B. 2001 Karyotype Analysis and Chromosome Banding Encyclopedia Of Life Sciences/ & 2001 Nature Publishing Group / www.els.net. Yuval G., I. T. Young and G. McNamara (2006) Spectral Imaging: Principles and Applications. Cytometry Part A 69: 735–747.

×