Talk:20.109(F09): Mod 2 Day 6 Readouts of DNA, Protein

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Contents

Information about genetic screen

T/R lab

INFORMATION TO BE FILLED IN BY EACH GROUP

team color library screened (K+ or P+) Approx # of colonies examined
(i.e., the # of colonies you looked at to find your 2 candidates)
Seq data b-gal data Notes
Red P+ Mutant 2 carries two mutations: A553I and H557A
  • Mutant 1 -- Mutant 2 -- 366+HA -- 366-HA
  • Light
  • 65 -- 66 -- 201 -- 232
  • Dark
  • 53 -- 25 -- 170 -- 1340
* Borrowed Mutant 2 from Green candidate 2
Orange K+ 2 Mutant 1 did not generate sequence data. Mutant 2 carries two mutations: G554V, V555S, S556A

WT without HA

  Light
    Average= 567
  Dark	
    Average= 961

WT-HA Tagged

  Light
    3 replicates: 221.9531881 -- 212.4645892 -- 233.3603896
    Average= 222.592722
    STDEV= 10.46257002
  Dark	
    3 replicates: 1588.465298 -- 174.3015663 -- 168.576792
    Average= 643.7812188
    STDEV= 818.1254186

Mutant 2

  Light
    3 replicates: 521.467 -- 526.889 -- 507.7317
    STDEV= 9.87505632370193
    Average= 518.696264320771
  Dark 
    3 replicates: 684.342 -- 668.203 -- 684.615
    STDEV= 9.39776470423833
    Average= 679.053216786013
Yellow P+ 2
Green
Blue
Pink K+ 2
Purple
Silver

All mutant candidates were digested with a cocktail of Nde and Mlu in 1X NEB3 for 45 minutes at 37° then loaded on a 1% agarose gel (1XTAE)

T/R Red, Orange, Yellow, Green
T/R Red, Orange, Yellow, Green
Lane Team Sample
1 1Kb DNA Ladder
2 Red candidate 1 Nde/Mlu digest
3 candidate 2 Nde/Mlu digest
4 blank
5 1Kb DNA Ladder
6 Orange candidate 1 Nde/Mlu digest
7 candidate 2 Nde/Mlu digest
8 blank
9 1Kb DNA Ladder
10 Yellow candidate 1 Nde/Mlu digest
11 candidate 2 Nde/Mlu digest
12 1Kb DNA Ladder
13 Green candidate 1 Nde/Mlu digest
14 candidate 2 Nde/Mlu digest
T/R Blue, Pink, Purple, Control digests
T/R Blue, Pink, Purple, Control digests
Lane Team Sample
1 1Kb DNA Ladder
2 Blue candidate 1 Nde/Mlu digest
3 candidate 2 Nde/Mlu digest
4 1Kb DNA Ladder
5 Pink candidate 1 Nde/Mlu digest
6 candidate 2 Nde/Mlu digest
7 1Kb DNA Ladder
8 Purple candidate 1 Nde/Mlu digest
9 candidate 2 Nde/Mlu digest
10 blank
11 1Kb DNA Ladder
12 Control with stuffer fragment Nde/Mlu digest
13 Control without stuffer fragment Nde/Mlu digest

Here is your Western data. If you'd like to see the blots themselves, they'll be in the lab at the front bench.
The "?" after your team color are associated with teams who labeled their containers on the lids rather than the tubs. These blots were possibly mis-designated during the washing and scanning of these blots since the lids were separated from the tubs.
If you'd like to note which of the 2 preparations of primary antibody you used, you can do so in the label under the blot's image

W/F lab

Digest Sample info

Please fill in the info below for your team for others to refer to:

team color library screened (K+ or P+) Approx # of colonies examined Seq data b-gal data Notes
Red
Orange
Yellow
Green
Blue P+ 2 red group's Colonies
  • Mutant 1 Mutant 2 366
  • Light/n
  • 17.4 47.5 163.1
  • 18.3 54.4 157.1
  • Average 17.85 50.95 160.1
  • Stdev 0.636396103 4.87903679 4.242640687
  • Dark
  • 18.9 43.6 178.5
  • 18.6 44.7 190.6
  • Average 18.75 44.15 184.55
  • Stdev 0.212132034 0.777817459 8.555992052
Pink K+ 2 :( Mutant 1 Light (Avg): 144 Mutant 1 Dark (Avg): 143 Mutant 2 Light (Avg): 327 Mutant 2 Dark (Avg): 301
Purple Using Red Group's Data (sequencing failed) N/A
Silver


Digest Images

Candidates were digested for 55' at 37 °C and run on a 1% agarose gel. Each gel was photographed at two different exposure times - use whichever one makes your DNA easier to see. Also see the second T/R gel for how control DNA (with and without stuffer fragment) runs.

Gel 1 Lane Gel 1 Sample Gel 1 Lane Gel 1Sample
1 1 Kb marker 6 Green Cand 1
2 Red Cand 1 7 Green Cand 2
3 Red Cand 2 8
4 9
5 1 Kb marker 10


Gel 2 Lane Gel 2 Sample Gel 2 Lane Gel 2Sample
1 1 Kb marker 6 Pink Cand 1
2 Blue Cand 1 7 1 Kb marker
3 Blue Cand 2 8 Purple Cand 1
4 1 Kb marker 9 Purple Cand 2
5 Pink Cand 1 10
Blue, Pink, Purple lower exposure
Blue, Pink, Purple lower exposure
Blue, Pink, Purple higher exposure
Blue, Pink, Purple higher exposure
Red, Green lower exposure
Red, Green lower exposure
Red, Green higher exposure
Red, Green higher exposure


Western images

Below are your Westerns. (Please crop out your image for use in the research article.) Some information may be lost in the digital images, so feel free to look at the original blots after journal club to get a good sense of where your bands are.

Westerns
Westerns
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