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Biotechnology > Embryo Transfer Technology > Flashcards

Embryo Transfer Technology Flashcards

1
Q

The 1st successful transfer of fertilized egg was reported in which animal and by whome?

A

Rabbit by walter and heape at cambridge 1891

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2
Q

1st success in which farm animals

A

Sheep and goat 1934

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3
Q

1st offspring after transfer of embryos on cattle and wine was in

A

1951

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4
Q

Enlist diff steps involved in embryo transfer

A 
Selection of donors
Synchronisation of oestrus
Induction of superovulation 
Collection of embryo
Evaluation of embryo
Selection of recepient 
Transfer of embryos
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5
Q

Embryo transfer techniques have been used in which animals

A

Extensively in cattle and more sporadically on sheep goat and swine.

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6
Q

Cow is preferable than hiefer as embryo donor T or F

A

T. Embryo transfer programme is based on milk yield or genetic superiority

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7
Q

What is criteria for selection of donor

A

The age of donor should be between 2 to 5 years
Free from generic diseases and conformational abnormalities
Exhibit refular oestrus cycle
Superior production traits
Record of sound reproductive performance Including successive fertility and AI

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8
Q

Enlist harmones used for superovulation

A

PMSG pregnant mare serum gonatotrophins
FSH Follicle stimulating harmone.
HMG human menopausal gonadotrophins

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9
Q

Stage of collection of harmones

A

In the early follicular or luteal phase of the oestrus cycle

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10
Q

Most frequently used harmones

A

PMSG and FSH

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11
Q

Doses of PMSG and FSH

A

PMSG one single injection 2000 -3000 IU

FSH 35-50 mg twice daily for 4 days

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12
Q

___________ and _______ after beginning the gonadotrophin treatment _______ is administered to induce oestrus . Insemination is preferred at ____ days after oestrus

A

48 and 60 hours
Prostaglandin (PG)
2days later the oestrus

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13
Q

Embryos are recovered _____ days after insemination

A

8days

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14
Q

In case of catlle embryo recovery is done at

A 
21 days 
10th day Pmsg and fsh
11th day fsh
12th day pg and pg/fsh
14th day anti pmsg oestrus and inseminations
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15
Q

In bovine embryo is collected by which method

A

Non surgical method

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16
Q

Explain non surgical method in bovines

A

Special catheter ie foleys catheter are introduced in the cervix into uterine horn and embryos are flushed with 250-350 ml flushing medium

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17
Q

In other farm animals like sheep goat pig which method is used

A

Surgical method is used

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18
Q

Classify embryos according to there morphological appearance

A
  1. Excellent embryos app dev stage and perfect morphology
  2. Good embryos app stage and slight deviation in morphology eg minor damage of zona pellucida and excluded blastomeres
  3. degenerated or retarted embryo inapp stage and major morph deviation
  4. Unfertilised ova
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19
Q

In bovines _______ and ______ are best suited as reciepients

A

Heifers and young cows

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20
Q

Transfer of embryo

A

Non surgical methods in cattle

Transfer the embryo in to the tip of the uterine horn without damaging the endometrium

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21
Q

For successful embryo transfer the most imp is __________

A

Embryo control

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22
Q

It is imp to transfer the embryo _______ position

A

Ipsilateral means into the horn bearing corpus luteum

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23
Q

Cryopreservation preserving them for long distance shipments

A

Optimal method for bovine embryo includes one step addition of 1.4M glycerol as cryoprotectant ,a 20 minutes equilibration period and 0.25ml straws as embryo containers , slow cooling down at -35 Celsius and subsequent plunging to liquid nitrogen -196 celsius . Embryo are thawed by placing the straws directly into warm water .

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24
Q

Application of embryo transfer

A 
Exploitation of femal repro system
Significant facilitation of import and export for valuable genetic material
Dev of new breeding concepts
Gene conservation by freezing techniques 
Twin production 
Intro of. New genes into closed herds
Manipulation of embryos
Gene transfer
25
Q

Enlist steps involved in invitro fertilisation

A

Oocyte recovery
Oocyte maturation
In vitro fertilisation of matured oocyte

26
Q

IVF is quite successful resulting in about ________ of fertilized eggs

A

70%-80%

27
Q

Which oocyte is selected for fertilization

A

2nd follicular phase of oestrous cycle …. Graffian follicles

28
Q

Size of preovulatory follicles of diff animals lying against the ovary wall

A

Cattle 15mm

Sheep pig 8mm

29
Q

_________ surgery can be used to recover oocytes from these follicles

A

Laparoscopy

30
Q

Oocytes are obtained from

A

Oviducts , follicles , and surface of ovary after ovulation

Immature ovary collected from ovary

31
Q

Culturing of oocytes

A

Cows are collected from the slaughter house and maintained in phosphate buffered saline at 30-32celsius for transportation

32
Q

Oocytes are washed twice with

A

HEPES and once with BMOC-3 branchat and olivhant medium for oocyte collection
After washing 5-10 oocytes are transferred to 0.2ml of BMOC drops under parrafin oil at 37 celsius with 5% co2 in 95% air for 12-32hrs

33
Q

_____ spermatazoa is essential for in vitro fertilisation

A

Capcitated

34
Q

Capcitation of spermatazoa

A

Reexposure to of spermatozoa to seminal vesicle

35
Q

In vitro fertilisation is done by

A

Oocytes are mixed with capcitated sperms in a petridish and incubated at 37 celsius with 5% CO2

36
Q

In IVF which stage of fertilized zygote is added to recipient

A

Morula stage 16 celled

37
Q

Sexing of sperm is done by

A

Flow cytometer attached with a sorter .

38
Q

Explain flow cytometer Method

A

Sperm is stained with hoechst 33342 and sex specific sperms are separated by the use of flow cytometer

39
Q

Method of flow cytometer has an accuracy of ________ of sexing sperms amd commercialy used in __________

A

90% cattle

40
Q

Methods of embryo sexing

A
  1. Y specific DNA probes presence or absence of Y DNA chromosome
  2. PCR amplify y chromosome specific repeat sequence DYZI 500-8000 times on y chromosome
  3. PCR-RFLP PCR based genotyping a pseudoautosomal region present between Xand Y these regions are called ZFX and ZFY
41
Q

Who described technique of micromanipulation

A

Lin injecting a mouse egg

42
Q

Define micromanipulation

A

It is a technique whereby sperm egg and embryo can be handled on inverted microscope via joysticks that hydraullicaly operate glass microtools

43
Q

1st to inject a sperm in oocyte by

A

Uehara and yanagimachi using hamster as a model

44
Q

Monoclonal antibody was 1st showed by

A

Kholar and milstein hybrids between myeloma tumor cells and antigen stimulating b lymphocytes

45
Q

Hybrid cells are formed by fusing the ________ cells and the __________ using a chemical __________

A

Myeloma cells
Antibody secreting B lymphocytes
PEG polyethylene glycol

46
Q

Hybrid cells have character of both parents ________ of the cancerous myeloma cells and _______ propert of B lymphocytes

A

Infinite growth

Antibody secreting

47
Q

Steps in production of monoclonal antibody production

A 
Immunization 
Fusion 
Selection of hybrid cells 
Cloning of hybrid cells
Large scale prod of ma cells
48
Q

Animal used for inoculation is

A

BALB/mice

49
Q

Cancerous myeloma cells which strain

A

Sp2/o

50
Q

Fused cells are grown on ________ medium

A

HAT hypoxanthine aminopterin thymidine

51
Q

The selection of hybrid cell formed is achieved by growing the cell on

A

HAT medium

52
Q

Cloning of hybrid cell is done by

A

ELISA

53
Q

For large scale production hybrido,as can be grown on tissue culture where they secrete upto ________ or in vivo as tumors in peritoneal cavity of balb mice where they produce upto _______

A

100ug 10-50ug/ml

40mg 2-20 mg/ml

54
Q

Methods of dna fingerprinting

A

RFLP restriction fragment length polymorphism
PCR
AFLP amplified fragment length polymorphism
STR short tandem repeats

55
Q

Dna fingerprinting syn

A

Dna profiling dna testing it is a process which uses fragments of dna to identify the unique genetic ,makeup of an individual

56
Q

In 1984, ___________ at university of leicester in England was able to distinguish diff among individuals based solely on their dna composition

A

Sir Alec jeffreys

57
Q

Read all methods from

A

Notes thanku dafffaaa ho ja amrinder

58
Q

App 12 hour after the end of normal heat a single non fertilized ova is released T or F

A

T

Biotechnology (4 decks)

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