Fragment sizes of H-Lo ladder
This presentation is the property of its rightful owner.
Sponsored Links
1 / 6

Fragment sizes of H-Lo ladder PowerPoint PPT Presentation


  • 45 Views
  • Uploaded on
  • Presentation posted in: General

Fragment sizes of H-Lo ladder. Gel 1: Pilar & Jim. 1 2 3 4 L 5 6 7 8. High Molecular Weight DNA (extraction). Origin (wells). That’s how it ideally should look like: nice single band.

Download Presentation

Fragment sizes of H-Lo ladder

An Image/Link below is provided (as is) to download presentation

Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author.While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server.


- - - - - - - - - - - - - - - - - - - - - - - - - - E N D - - - - - - - - - - - - - - - - - - - - - - - - - -

Presentation Transcript


Fragment sizes of h lo ladder

Fragment sizes of H-Lo ladder


Fragment sizes of h lo ladder

Gel 1: Pilar & Jim

1 2 3 4 L 5 6 7 8

High Molecular Weight DNA

(extraction)

Origin (wells)

That’s how it ideally should look like: nice single band

Here there is some HMW DNA, but also a lot of degraded DNA. This probably would still be ok for PCR

This is RNA

PCR product

Probably overloaded (too much DNA). Could also be too much template DNA in the PCR.

HMW DNA (top)

1 (Pilar)-Sockeye #10-Salt

2 (Pilar)-Coho #2-Salt

3 (Pilar)-Sockey #10-Qiagen

4(Pilar)-Coho #2-Qiagen

L-Ladder

5-(Jim)-Sockeye #10-Salt

6-(Jim)-Chinook #4-Salt

7-(Jim)-Sockeye #10-Qiagen

8-(Jim)-Chinook #4-Qiagen

PCR product (bottom)

1-(Jim)-Chinook #4-Salt

2-(jim)-Chinook #4-Qiagen

3(Jim)-Sockeye #10-Salt

4-(Jim)-Sockeye #10-Qiagen

L-Ladder

5-(Pilar)-Sockeye #10-Qiagen

6-(Pilar)-Coho #2-Qiagen

7-(Pilar)-Sockeye #10-Salt

8-Negative control

Conclusion:

No obvious effect of salt vs Qiagen extraction, either in HMW or PCR product

1 2 3 4 L 5 6 7 8


Fragment sizes of h lo ladder

Gel 2: Claudia, Kristi & Nick

L 1 2 3 4 5 6 7 8

Good HMW DNA with little degradation, but the PCR didn’t work well


Fragment sizes of h lo ladder

L 1 2 3 4 5 6 7 8

Gel 3: Brian & Gang

Weak HMW DNA with some degradation, but the PCR worked well in three samples (all Quiagen extractions!)

HMW DNA

L: ladder

1: DNA A Quiagen (GX)

2: DNA B Quaigen (GX)

3: Chinook 2 salt (GX)

4: Sockeye 7 salt (GX)

5: Coho 3 salt (BK)

6: sockeye 8 salt (BK)

7: Coho 3 Quiagen (BK)

8: Sockeye 8 Quiagen (BK)

PCR product

L: ladder

1: Sockeye 7 salt (GX)

2: Coho 3 salt (BK)

3: Sockeye 8 Quiagen (BK)

4: Coho 3 Quiagen (BK)

5: DNA A Quiagen (GX)

6: DNA B Quagen (GX)

7: Chinook 2 salt (GX)

8: control

L 1 2 3 4 5


Fragment sizes of h lo ladder

Gel 4: Alfred and Danny

This is probably template DNA, i.e. there was too much in the PCR reaction – it still worked though

Good PCR products in 5 of 7 samples

Good HMW DNA in all samples (#8 a bit weak)

Degraded DNA

RNA


Fragment sizes of h lo ladder

Gel 4: Bill & Thomas

Gel broke to two pieces

DNA looks ok in 4 samples

Something strange going on here – you can’t see it on the picture, but on the gel the stain was only on the bottom surface of the gel – a lesson that you can’t see everything on a picture.

PCR worked fine as well – this is actually the gel where we saw most straight away.

DNA

1.Chinook 1 salt

2.Chinook 1 Qiagen

3. Sockeye 6 Qiagen

4. Steelhead 3 Qiagen

5. Sockeye 9 Qiagen

6. Steelhead 3 salt

7. Sockeye 9 salt

8. Ladder

PCR product

Ladder

1. Sockeye 9 salt

2.Steelhead 3 salt

3. Sockeye 9 Qiagen

4. Steelhead 3 Qiagen

5. Sockeye 6 Qiagen

6. Chinook 1 Qiagen

7. Negative control

8. Chinook 1 salt


  • Login