Professional Documents
Culture Documents
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10 20 .
- 13
) Balances ( 3 4
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- 14
- 15
- 16
- 17
Refregerator .
- 18
Deep Freezer
.
- 19
Steamer .
- 20
pH meter pH
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Forceps
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Forceps .
- 30
Needels .
- 31
Scalpels .13 - 12 - 11 - 10
- 32
Binocular
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- 33
Air Condition
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- 34
Shaker ) (
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- 35
Centerfuge .
- 36
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) ( . Lightmicroscope
- 38
- 39
- 40
)( ELISA test .
- 41
Reader .Microplate
- 42
10
- 43
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Timer 1000 10000
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- 44
Raks
12 24 36.
- 45
Water bath
.
- 46
Gloves
.
- 47
Over head .
- 48
Mask
.
- 49
Over shoes
.
- 50
Lux meter
.
- 51
.Chemicals .
- 52
- 53
Filter papers .
- Media preparation
-1 .
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- General equipment
-1 .
-2 .
-3 .
-4 .
-5 .
-6 .
-7
.
- Growth room :
-1 27 17.
-2 .
-3 .
-4 .
-5 .
-6 .
-7 .
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12
: ) (
Media
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- 1 Media Composition
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gelling agent
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Murashige and Skoog
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N6 Chu
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E1
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400
250
2100
600
450
-
185
400
2830
166
463
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MS B5 Nitschs
185
68
950
720
-
250
150
2500
150
134
370
170
1900
1650
440
-
13
White
750
19
80
-
MgSO4. 7H2O
KH2PO4
NaH2PO4. H2O
KNO3
NH4NO3
CaCl2. 2H2O
(NH4)2 SO4
3
10
2
0.25
0.025
0.025
0.80
43
1.6
4.4
3.3
1.5
0.80
27.8
37.8
-
25
10
0.25
0.025
0.025
27.8
37.8
-
3
10
2
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0.025
0.75
43
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22.3
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27.8
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25
25
20
20
30
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100
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10
1
1
100
5.5
0.5
0.5
0.5
100
2
5.8
0.01
0.01
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3
5.8
H3BO3
MnSO4. 4H2O
MnSO4. H2O
ZnSO4. 7H20
NaMoO4. 2H2O
CuSO4. 5H2O
CoCl2. 6H2O
KI
FeSO4. 7H2O
Na EDTA. 2H2O
EDTA Na ferric salt
)Sucrose (g
Thiamine HCl
Pyridoxine HCl
Nicotinic acid
Myo-inositol
Glycine
Folic acid
Biotin
pH
:
) White: White (1953) Am. J. Bot. 40, 517 524). MS: Murashige and Skoog (1962
Physiol. Plant. 15 : 473). B5: Gamborg et al.( 1968) Exp. Cell Res. 50: 151). Nitschs:
(Nitsch and Nitsch 1969 ) Sci., N. Y., 163: 85).N6: Chu (1978); Pro. Symp. Plant
Tissue Culture, Sci. Press, Peking P. 43). E1: Gamborg et al. (1983). Plant Cell Rep. 2:
209).
-:Growth Regulators
.
.
) ( .
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-1 Auxins
indole-3 acetic acid ) (IAA indole 3
butyric acid ) (IBA 4 2 )D) 2,4 4 ,2
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(NAA IAA
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to 10 mg /L 0.1
14
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100 1 = 1.
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IAA DNA
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(P chlorophenoxy acetic acid (PCPA
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5 4 2
(Trichlorophenoxy acetic acid (2, 4,5 T 2,4,5
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2 4
2 methy 4 chlorophenoxy acetic acid (MCPA).
-4
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3,6 dichloro 2 methoxy benzoic acid. (dicamba).
15
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Cytokinins
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6(4hydroxy3methyltrans-2 butanylamino) purine
)Zeatin
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to 10 mg/L 0.01
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(El-Shobaky (1991 2 -2/1
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pH pH pH meter :
pH 4.5 7 .
pH 4.5 :
- 1 IAA .
- 2 .
- 3 .
- 4 .
pH ) ( 8 :
- 1 .
- 2 .
- 3 .
- 4 .
- 5 .
- 6 .
pH pH 5 3
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Media types
17
- 1
Solid media
7 2 / 2 /
pH 5.7 45
.
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Liquid media
pH 5.5
Shaker )(
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Semi-solid media
Solidifying Agents
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Gelrite,
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Media Preparation
.
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1
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18
.
) (Stocks
.
Stocks
100 )
( :
Micro nutrients
100 X
1X
mg/100L
mg/L
83
0.83
KI
620
6.2
H3BO3
860
8.6
ZnSO4.7H2O
2230
22.3
MnSO4.4H2O
25
0.25
NaMoO4.2H2O
2.5
0.025
CuSO4.5H2O
2.5
0.025
CoCl2.6H2O
3730
37.3
Na2.EDTA
2780
27.8
FeSO4.7H2O
FeSO4. 7H2O
Na EDTA. 2H2O 45
100.
85 100.
70 100..
1
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20
)MS) Stocks and Media Preparation
Stocks ) ( - 4
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4
5
6
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-2 Selective of an explant
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Explant
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Age of Explant
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.
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Size of Explant
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Season of Explant
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Growth Chamber
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- Light requirement :
- 1
Light intensity
Initiation Morphogenetic processes
1000 3000
10000 .
).(El-Shobaky 1991
- 2
Photoperiod
16 8 12
.
-
Heat requirement
27 - 22
35 - 27
18 . 25-20
.(El-Shobaky (1991
21
: :
Acclimatization
10000
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23
:
Production of Banana (Mousa sp.) through tissue culture
.
58 18
) Banana Bunch Top Virus Banana
(Mosaic
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- 11 2 27
1500 16 .
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: Multiplication stage
:
MS salt+5mg/LBAP + 30 g/L Sucrose + 100 mg/L Inositol + 2g/L Gelrite or 8 g/L Agar
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MS salt+1mg/LBAP + 30 g/L Sucrose + 100 mg/L Inositol + 2g/L Gelrite or or 8 g/L Agar
: Rooting stage
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) ( . :
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