Talk:20.109(F09): Mod 1 Day 3 Agarose gel electrophoresis

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Here is an example of a gel with all the expected bands, as well as a gel with the needed bands excised.

You can use these in your lab report if you so choose.

FragsMod1.jpg
Excisedfrag.jpg


T/R lab

Red Orange Yellow Green
TR Red.BMP
TR Orange.BMP
TR Yellow.BMP
TR Green.BMP
Blue Purple Pink
TR Blue.BMP
TR Purple.BMP
TR Pink.BMP

And here's the gel with your recovered fragments/backbones, ready for ligation.



TR Blue, Violet, Pink.BMP


Lane Sample Lane Sample
1 Blue Frag 6 Pink BKB
2 Blue BKB 7 --
3 Purple Frag 8 --
4 Purple BKB 9 --
5 Pink Frag 10 ladder








Lane Sample Lane Sample
1 Red Frag 6 Yellow BKB
2 Red BKB 7 Green Frag
3 Orange Frag 8 Green BKB
4 Orange BKB 9 --
5 Yellow Frag 10 ladder
TR Red,Orange,Yellow,Green.BMP








Red Team: Yours didn't show up well. Yours was the first sample I loaded and the gel was cracked so the sample went through the bottom of the gel. I tried to pipette as much of the sample as possible out of the broken gel and into this gel, but I didn't get much. Assume that your sample is about like that of Orange to do your HW due on Thursday.


Red Team: I re-ran 5 ul of your product with 10 ul water and 2 ul loading dye. I ran it for 20 min at 125V.

Please use this image for your final lab report.

Re-run Red TR.BMP
Lane Sample
1 10 ul ladder
2 5 ul backbone + 10 ul H20 + 2 ul loading dye
3 --
4 5 ul fragment + 10 ul H20 + 2 ul loading dye



W/F Lab

Red Green Blue
WF Red.BMP
WF Green.BMP
WF Blue.BMP
Purple Pink Pink Post-stained fragment part
WF Purple.BMP
WF Pink.BMP
WF Pink post stain 30 min-no frag.BMP

Green Group: For some reason, your ladder did not show up properly. Please use the example images at the top of the page for your calculations for next time.

Pink Group: Unfortunately, it appears that you didn't have a PCR product. We had discussed possibilities of why we didn't see a band at first, namely the EtBr migrating out of the gel. This was a good hypothesis since we couldn't see the last few bands of the ladder. While you were recovering the backbone, I post-stained the gel in an EtBr / TAE solution for 30 min followed by two rinses in DI water. I cut through the ladder, but we can now see the 500 bp band, but still can't see a fragment. Since we can see the 500 bp band, it means that your band was not run off of the gel. You will use fragment from the Purple group, so use that image for your calculations for next time.

And here's the gel with your recovered fragments/backbones, ready for ligation.



WF check gel.BMP


Lane Sample Lane Sample
1 ladder 8 Blue BKB
2 Red BKB 9 Blue Frag
3 Red Frag 10 Pink BKB
4 Green BKB 11 Purple BKB
5 Green Frag 12 Purple Frag
6 -- 13 --
7 ladder 14 ladder