RNA-B EcR knockdown

Submitted by Anonymous (not verified) on 25 April, 2013 - 09:12

Phenotype CV:

General

Experiment Batch ID:

RNAB

Start Date:

19 March, 2012 - 09:06

End Date:

20 April, 2012 - 00:00

Experiment Description:

Knockdown of EcR. CPY185 used as control

Sample

Organism:

Lepeophtheirus salmonis

Sex:

female

Number of Individuals (Start):

39

Developmental Stage CV:

PreAdult

Fragment description

Fragment length:

667bp

Fragment concentration:

600ng/ul

Fragment Label ID:

18

Primer Group:

Validation

Efficacy of Knock Down:

F>20%

Phenotype

Number of Individuals (End):

14

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F18 treated lice

F18 treated lice close up

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Sampled lice had no eggstrings except one whioch most likely have not been injected properly.

K F81

Submitted by Sussie Dalvin on 23 April, 2013 - 13:27

Phenotype CV:

female reproduction

General

Experiment Batch ID:

K

Experiment Contact:

Sussie Dalvin

Start Date:

3 April, 2013 - 10:46

End Date:

13 May, 2013 - 00:00

Sample

Organism:

Lepeophtheirus salmonis

Sex:

female

Number of Individuals (Start):

34

Developmental Stage CV:

Adult

Target description

Primary target ID:

EMLSAT00000006465, EMLSAT00000006465-702312 (mRNA) Lepeophtheirus salmonis

Gene or target symbol:

Etif_g1565

Fragment description

Fragment length:

485bp

Fragment concentration:

600ng/ul

Sense sequence:

taccgactttgaaatcaagcatccgttagaacatgaatggactctttggtattataatcatgatccacaaaatgcatgggaagacaatctcaaagaggtaatttcattcaatactgtggaagatttttgggccatatataatcatatcgagttagcctccgagataagtgataaatgtgactattgtctctttaaaaagggagtaaaacccatgtgggaagacgcttctaattgtaatggaggacgctgggtttatagtactcctgtaaatagaagaaaatcatttgacggaggactagatgtgaattggctagaagtgatgatgtctttaataggtgaagcttttgaagggtctggtgatttgattaatggtgctgttgttagtcttcgtaataaatcagataaaatatcaatttggttgggtacagcgtatgacgaggatgacggcatccgaagaattggagcgattctcaagaaaagacttg

Fragment Label ID:

F81

Primer Group:

b1434

Fragment Producer:

licelab

Fragment Producer Contact:

Heidi Kongshaug

Validation

Efficacy of Knock Down:

F>30%

Phenotype

Number of Individuals (End):

20

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late hatching + 5 animals where one or both eggstrings did not hatch, one very late hatch (+14 days)

K F94

Submitted by Sussie Dalvin on 22 April, 2013 - 14:55

Phenotype CV:

female reproduction

General

Experiment Batch ID:

K

Experiment Contact:

Sussie Dalvin

Start Date:

14 April, 2013 - 14:54

End Date:

13 May, 2013 - 00:00

Sample

Organism:

Lepeophtheirus salmonis

Sex:

female

Number of Individuals (Start):

36

Developmental Stage CV:

PreAdult2

Adult

Target description

Primary target ID:

EMLSAT00000009747, EMLSAT00000009747-705594 (mRNA) Lepeophtheirus salmonis

Gene or target symbol:

UN15_g1178

Fragment description

Fragment length:

445bp

Fragment concentration:

600ng/ul

Sense sequence:

Aaaaggataaggaaccctcaccaccaagtaaaaaggcctccactacgagtctaaagaaaaaaacactagagcgacaaaaatccaagaaggaagagactgacaaagcagagaatgaggatggaaaaaactccataaaaagaaaggacacattccgacggcgattttcacttcgaatgaggaaaacagctttaaagcactcactttcatctgcaacaacttcaactgcaacaagcagcgaatatcaatccgatgatggaggagagtctgatgactattccgacgacgacgacgacgatgatgatgaggacactctcgtaaatagtatcaaactcagctcacagactttatcccttgggggagcagatgatcacttttctgataaggatggctcagcgcatcgactcacagaactcttcatcaacgacatcaatcttattgtggataaaactcttgctcttcctaatggcccagataaaattaagatgatccattctgttatagaatccattaaacccttgtacaacatggcgacactcaatcttca

Fragment Label ID:

F94

Primer Group:

b1473

Fragment Producer:

licelab

Fragment Producer Contact:

Heidi Kongshaug

Validation

Efficacy of Knock Down:

F>70%

Phenotype

Number of Individuals (End):

10

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F94(UN15_g1178): Funny looking and short eggstrings in many individuals. Hatched copepodites looked normal

Sectioned animal RFK 13: ovaries very abnormal. Malformed eggs in GS, necrosis in some subcuticular tissue mailnly posterior in the cephalothorax. One type of gland like structures stain green, other glands appear normal.

The student Julia validated the RNAi lice with qPCR 130916

5' and 3' race have been performed

K F77

Submitted by Sussie Dalvin on 22 April, 2013 - 14:51

Phenotype CV:

female reproduction

General

Experiment Batch ID:

K

Experiment Contact:

Sussie Dalvin

Start Date:

3 April, 2013 - 14:49

End Date:

13 May, 2013 - 00:00

Sample

Organism:

Lepeophtheirus salmonis

Sex:

female

Number of Individuals (Start):

36

Developmental Stage CV:

PreAdult2

Adult

Target description

Primary target ID:

EMLSAT00000003661, EMLSAT00000003661-699508 (mRNA) Lepeophtheirus salmonis

Gene or target symbol:

UN8_g528

Fragment description

Fragment length:

507bp

Fragment concentration:

600ng/ul

Antisense sequence:

Aggcagagccaatgtgtctccatcaaaaaaaaagaagggcaatgaaaagaaaaagattgaaaaacatgtgcaaaacgggagtcgactctatcatcatcaacaaaccaaaaagcaccccaaagttcgccaacgtatcaactctggagaagggagttccgaagcggattccttatcatgggatgactacgatgttgctgctcacaaaaggagacagggaatacatacccttgagaacgttgagtctctcatctcaacttattcctctccttccctaagttcttcttccgcctcatccaaggagtccactccagagatgcaccaggaaggaaaacttttacttcaggaaatgaaggaactcagaaaggagattaagaaaatgggagatacttctgtccttgagaacaacactcactccaaggtgatacaactacttccaacggactattcaattgaggaactccaagaaatgatgaacaaaatggaggaagaggatcgttgttgggattc

Fragment Label ID:

F77

Primer Group:

b1400

Fragment Producer:

licelab

Fragment Producer Contact:

Heidi Kongshaug

Validation

Efficacy of Knock Down:

F>90%

Phenotype

Number of Individuals (End):

19

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F77 (UN8_g528): 12/21 animals did not carry eggstrings. Hatched copepodites looked normal.

RFK8 sectioned, looked fairly normal but dorsally the immature (posterior) part of the ovary looks abnormal. Egg anterior in GS (about to be vitellogenised) show abnormalities

The intestine seem to stain patchy especially in the anterior part (significant ?).

Subcuticular tissue appears normal

Some animals produced normal copepodites. It can be expected that all normal eggproduction would have ceased within one or 2 cycles.

H F60

Submitted by Sussie Dalvin on 9 April, 2013 - 13:26

Phenotype CV:

General

Experiment Batch ID:

H

Experiment Contact:

Sussie Dalvin

Start Date:

15 January, 2013 - 13:17

End Date:

25 February, 2013 - 00:00

Sample

Organism:

Lepeophtheirus salmonis

Sex:

female

Number of Individuals (Start):

30

Developmental Stage CV:

PreAdult2

Adult female

Target description

Primary target ID:

EMLSAG00000003343, EMLSAG00000003343-686109 (gene) Lepeophtheirus salmonis

Gene or target symbol:

FK506

Fragment description

Fragment length:

542bp

Fragment concentration:

600ng/ul

Sense sequence:

ccacgggcccaggaactgttcatttgacaggttatttgattcccgacactgacgacatgattgacgacctcgattcagagtctgaatgtttnnnnnnnnnnnnnnnnnnagctcctgtccttgttaatggaaagggcaagaacaccaaacgtaagcctgataatagcttaattgatactcctaataaaaaagctaaaaaagagttggaacttgctagagctaaaaaagttcagtccgatcttaacttgaagaagaagagtcaaaagaaagtcgagtcttcggatgatgataacgatatggaagcagatgatgatacgtcggatgatggaatattagatttcactgagaattacgctgaagaagaggactctgtcgatgaatctgatgacgaatacgaggatgattcttcagtagaagaaggcgattcctctccaacaaagaaaattctcgagaaaatagccaataaaaaggagtcttctctcactactcccaagaacaataagaaaaaggataatgcctccactcctaaaggcgctactccc

Fragment Label ID:

F60

Primer Group:

FK506_f_b1313

Validation

Efficacy of Knock Down:

F>14%

Phenotype

Number of Individuals (End):

13

Phenotype:

no reproduction

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Females looked normal, but none had produced any eggstrings. Some animals had small empty eggstrings hanging behind them. Some eggs appeared vitellogenised but thin. Digestion looked normal.

Little pathology on RNAi animals sections. Eggs not vitellogenized

Uncertain about number of animals at endpoint

E F42

Submitted by Heidi Kongshaug on 12 March, 2013 - 10:38

Phenotype CV:

General

Experiment Batch ID:

E

Experiment Contact:

Frank Nilsen

Start Date:

15 October, 2012 - 10:19

End Date:

26 November, 2012 - 00:00

Sample

Organism:

Lepeophtheirus salmonis

Sex:

female

Number of Individuals (Start):

39

Developmental Stage CV:

PreAdult2

Adult

Target description

Primary target ID:

EMLSAT00000005071, EMLSAT00000005071-700918 (mRNA) Lepeophtheirus salmonis

Gene or target symbol:

g13677_Rheb

Fragment description

Fragment length:

460bp

Fragment concentration:

600ng/ul

Sense sequence:

atggggcctcccaagtctcgtaaaattgccttaatgggagctgtttccgtgggcaaatcttctctggccatccaatttgtacaaggacaatttgtcgattcctacgaaccaacaatcgaaaatacatttaccaaaacagttaaaattaatagacaagaatatgaaatttgtcttgttgatacagcaggccaaaatgaatactctatctttccattggaatattctgtaggaatagatggatatgttcttgtatactccattgattcacctaaaagcttagaagtttgtcaaatcatacaggaaaaattaatagaattcacaggaaacccaaatgttcccattgtcttggttggcaataaaaatgatttgtatatggaaagatatgtttcctttgaagaaggcaaaagaagagctgaagaaatgaaagcagtgtttcttgagacctctgccaaggaaaacc

Fragment Label ID:

F42

Primer Group:

Rheb_f_b1063

Fragment Producer:

licelab

Fragment Producer Contact:

Heidi Kongshaug

Validation

Efficacy of Knock Down:

F>60%

Phenotype

Number of Individuals (End):

25

Phenotype:

Low reproduction and most are missing blood in the gut

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Eggestrenger:

12.11 F42 ser ut som der ikke er noen sunne eggstrenge mer (har vært tidligere).

15.11 Lite blod i F42.

E F34

Submitted by Christiane Trösse on 12 March, 2013 - 09:05

Phenotype CV:

General

Experiment Batch ID:

E

Experiment Contact:

Sussie Dalvin

Start Date:

15 October, 2012 - 00:04

End Date:

26 November, 2012 - 00:00

Experiment Description:

24 females recovered (62%), 7 with normal looking eggstrings, that hatch. Animals with eggstrings have blood in the gut, animals w/o eggstrings have little or no blood. Theis group of animals looked almost normal and was reproducing until 3 days before termination (last check). Seem to have been caught just when the system broke down. Reproducing animals had normally developed cops (morph and numbers)

Sample

Organism:

Lepeophtheirus salmonis

Sex:

female

Number of Individuals (Start):

39

Developmental Stage CV:

PreAdult2

Adult

Target description

Primary target ID:

EMLSAG00000003242, EMLSAG00000003242-686008 (gene) Lepeophtheirus salmonis

Gene or target symbol:

g7499_LsRaptor

Fragment description

Fragment length:

451bp

Fragment concentration:

600ng/ul

Sense sequence:

tcaagattcctcctcaactgccgctactccttggagactccttgaacgaatgaaaactgtgagtgttgctcttgtggtatgcctcaatgtcggggtggatcctccggatgtgacgaaaacctccccttgtgcttgtattgaggcatggattgatccaactcttctcaatccaacaagagcacttaatctcattggatctgctcttctaagacaatatgaacgttggcaacccagggctaggtatagacaatctttggacccaactgtggacgaagtccgacgcctagctatggctctacgtaaaaacgctagggatgaaagagttctttttcattataatggtcatggggtcccgaaaccaacggcaaacggagagatttgggtttttaatcggaattacactcagtatattcccctgagtctctacgatcttcaaacatggatgggctcc

Fragment Label ID:

F34

Primer Group:

Raptor_f_b918

Fragment Producer:

Heidi

Validation

Efficacy of Knock Down:

F>50%

Phenotype

Number of Individuals (End):

24

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Adult females recovered at termination of RNAi experiment. Experiment E.

D F29

Submitted by Heidi Kongshaug on 6 March, 2013 - 14:49

Phenotype CV:

female reproduction

General

Experiment Batch ID:

D

Experiment Contact:

Frank Nilsen

Start Date:

24 August, 2012 - 00:00

End Date:

3 October, 2012 - 00:00

Sample

Organism:

Lepeophtheirus salmonis

Sex:

female

Number of Individuals (Start):

36

Developmental Stage CV:

PreAdult2

Target description

Primary target ID:

EMLSAG00000006214, EMLSAG00000006214-688980 (gene) Lepeophtheirus salmonis

Gene or target symbol:

g2282_trehalose

Fragment description

Fragment length:

501bp

Fragment concentration:

600ng/ul

Sense sequence:

cttcaattgcatcacaagctgcgcttctcatgggctgtagtttgggttggtcaagccccgtaatcgtattattaagtaaatcaaaggatggatttgttctcagcaagtcagaagagggatggttggcctccatattattctttggtgccatagcaggaggtccttgcgctggatacctgattgacaaatctggtcgaaaattggcgttaggagtcaatgctgtattgtatgcattgtctcttggagttattgtatctgcgcaatataagtggatgctatttgcaggacgttttttaaatggtgtgtcatgtgggtttacttctattagtgccccaacttatatctcagaaatttcttcagcgagattgattgaaagttggcgttggattactgtttctattattataatggattccatatggttattttttctctattttattccggaatccccagtttatcatctgataaataacaacgaacctttagcaaggagcgttc

Fragment Label ID:

F29

Primer Group:

g2282_trehaloseF_b836

Fragment Producer:

licelab

Fragment Producer Contact:

Heidi Kongshaug

Validation

Efficacy of Knock Down:

F>59%

Phenotype

Number of Individuals (End):

12

Phenotype:

a bit short eggstrings (?), other wise normal. One very strange looking animal

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Test.

Did not find the sampling sheet for D experiment.

The Tif pictures were to big.

E F50 (Ferr_sub)

Submitted by Christiane Trösse on 21 February, 2013 - 11:54

Phenotype CV:

General

Experiment Batch ID:

E

Experiment Contact:

Christiane Trösse

Start Date:

15 October, 2012 - 00:00

End Date:

26 November, 2012 - 00:00

Experiment Description:

Ferr:_sub

Sample

Organism:

Lepeophtheirus salmonis

Sex:

female

Number of Individuals (Start):

38

Developmental Stage CV:

PreAdult2

Adult

Target description

Primary target ID:

EMLSAG00000010792, EMLSAG00000010792-693558 (gene) Lepeophtheirus salmonis

Gene or target symbol:

ferr_sub

Fragment description

Fragment length:

568bp

Fragment concentration:

600ng/ul

Sense sequence:

acaggagcccatggatcttgtaacataggttcacagacggacccaaaaggaaaagcaactcctgaagatccaacaaattatcatgaattttgtatttcttcattaaggaaccaagtcaaaatggaatttgaagcagctcttcaatatctagttatgggagcctactttgatcaagacactatcaatctcaagggatttggtaagttcttttgggaaagtgctgacgaagaaagacagcatggattacaatttatcaaatacttgcgacacagaggagacaagggtctcgatttctttcaaggagatattgagcctattctaagaaagtattcatggagtgatggacttgatgctcttcgtgacgctcttatgatggaaaaaaaagtttcgggttccataaagaagatcatagatatttgcagtgttgaaggaaagcaagattatctctctggagattggctcactggtacatggttggaagagcaaatggatggacaaagaaagttagcaggacttatcaacacgttaagcaactttagaattgaccatgaggctcttggtgactgga

Fragment Batch ID:

F50B1

Fragment Label ID:

50

Primer Group:

Ferr_sub_f_b632

Fragment Producer:

Christiane Trösse

Fragment Producer Contact:

Christiane Trösse

Validation

Efficacy of Knock Down:

F>85%

Phenotype

Number of Individuals (End):

18

Phenotype:

Most lice didn’t have egg strings at termination (but had first and second which had fallen off and were found in the filter). No reproduction.

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Average: 3.5 (1 vote)

7a: 6 of 9 lice have no or very short egg strings. 5 of 9 lice have very little gut content.

7b: 2 of 4 lice have no or very short egg strings. 2 of 4 lice have very little gut content.

7c: 3 of 5 lice have no or very short egg strings. 5 of 5 lice have very little gut content.