Chromatin experiment

Discussion of all aspects of biological molecules, biochemical processes and laboratory procedures in the field.

Moderators: honeev, Leonid, amiradm, BioTeam

Post Reply
2468
Garter
Garter
Posts: 23
Joined: Thu Nov 01, 2012 12:48 pm

Chromatin experiment

Post by 2468 » Thu Nov 01, 2012 12:57 pm

Hey,
I need help with this question...does anyone have any idea?
I've uploaded the results as a attachment.

Question: Consider the six lanes from the left (where the most left lane is lane 1 and lane 4 and 5 are empty). Match the correct explanation for the bands visible in the different lanes.

Explanations:
1. In this lane the DNA has been almost completely digested because the nucleosome had been extracted before nuclease treatment

2. The high molecular weight bands in this lane represent DNA protected from the nuclease

3. The strong low molecular band in this lane represents the DNA that stretches between nucleosomes

4. High molecular weight bands in this lane represent the undigested DNA

5. The bands in this lane represent the molecular weight standards

6. The band at approximately 200 bp in this lane represents the DNA that is wrapped around the nucleosomes

7. The lack of bands in this lane (except for a very faint band at at 200bp) demonstrates that EDTA has effectively inhibited the nuclease enzyme


Answers to match the explanations: Lane 1, Lane 2, Lane 3, Lane 6, Wrong Interpretation.


Thank you :)
Attachments
chromatin results pic.jpg
Results pic

User avatar
JackBean
Inland Taipan
Inland Taipan
Posts: 5694
Joined: Mon Sep 14, 2009 7:12 pm

Post by JackBean » Thu Nov 01, 2012 4:19 pm

So what's your idea?
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

2468
Garter
Garter
Posts: 23
Joined: Thu Nov 01, 2012 12:48 pm

Re:

Post by 2468 » Fri Nov 02, 2012 6:19 pm

JackBean wrote:So what's your idea?


Well so far i got : explanation 2 = lane 1
explanation 3 = lane 3
explanation 4 = lane 2
explanation 7 = lane 1

Not to sure about the rest...so need help with them! :)

User avatar
JackBean
Inland Taipan
Inland Taipan
Posts: 5694
Joined: Mon Sep 14, 2009 7:12 pm

Post by JackBean » Fri Nov 02, 2012 10:15 pm

how can you have two explanations for one lane?

Do you really think that the lane 3 has such strong band?
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

2468
Garter
Garter
Posts: 23
Joined: Thu Nov 01, 2012 12:48 pm

Re:

Post by 2468 » Fri Nov 02, 2012 10:27 pm

JackBean wrote:how can you have two explanations for one lane?

Do you really think that the lane 3 has such strong band?



Just to help you count...Theres 7 explanations and 5 options. Some answers MIGHT be a repeat... :)

Well, I wouldnt be asking for help if I knew the answers myself if that isnt obvious... :/

User avatar
JackBean
Inland Taipan
Inland Taipan
Posts: 5694
Joined: Mon Sep 14, 2009 7:12 pm

Re: Chromatin experiment

Post by JackBean » Sat Nov 03, 2012 2:40 pm

OR some of them may not be used ;) :roll:

Don't you think that explanations 2 and 7 are contradictory (high-molecular bands vs. lack of bands)?
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

Cat
King Cobra
King Cobra
Posts: 635
Joined: Thu Feb 14, 2008 7:40 pm

Post by Cat » Sat Nov 03, 2012 3:46 pm

I am not completely sure about lanes 1 and 2, but 100% sure about 3 and 6.

Lane 2 (smear starting in the middle of the gel ending with strong band of low molecular weight)= 1. In this lane the DNA has been almost completely digested because the nucleosome had been extracted before nuclease treatment

Lane 1 (smear starting at the well and ending in the middle of the gel means undigested DNA)= 4. High molecular weight bands in this lane represent the undigested DNA

Lane 6 = 5. The bands in this lane represent the molecular weight standards
Lane 3 = 7. The lack of bands in this lane (except for a very faint band at at 200bp) demonstrates that EDTA has effectively inhibited the nuclease enzyme

2468
Garter
Garter
Posts: 23
Joined: Thu Nov 01, 2012 12:48 pm

Re:

Post by 2468 » Sat Nov 03, 2012 4:32 pm

Cat wrote:I am not completely sure about lanes 1 and 2, but 100% sure about 3 and 6.

Lane 2 (smear starting in the middle of the gel ending with strong band of low molecular weight)= 1. In this lane the DNA has been almost completely digested because the nucleosome had been extracted before nuclease treatment

Lane 1 (smear starting at the well and ending in the middle of the gel means undigested DNA)= 4. High molecular weight bands in this lane represent the undigested DNA

Lane 6 = 5. The bands in this lane represent the molecular weight standards
Lane 3 = 7. The lack of bands in this lane (except for a very faint band at at 200bp) demonstrates that EDTA has effectively inhibited the nuclease enzyme



Thank you! :)

User avatar
JackBean
Inland Taipan
Inland Taipan
Posts: 5694
Joined: Mon Sep 14, 2009 7:12 pm

Re:

Post by JackBean » Sat Nov 03, 2012 6:02 pm

Cat wrote:Lane 6 = 5. The bands in this lane represent the molecular weight standards

Really? You see there distinct bands? Something in regular distances?
http://www.biolib.cz/en/main/

Cis or trans? That's what matters.

Cat
King Cobra
King Cobra
Posts: 635
Joined: Thu Feb 14, 2008 7:40 pm

Re: Re:

Post by Cat » Sun Nov 04, 2012 6:54 pm

JackBean wrote:
Cat wrote:Lane 6 = 5. The bands in this lane represent the molecular weight standards

Really? You see there distinct bands? Something in regular distances?


This is a bad picture of a bad gel as far as I can tell. And yes, I do see familiar pattern. Here is another example of poor gel:

http://aem.asm.org/content/66/4/1423/F1.expansion.html

2468
Garter
Garter
Posts: 23
Joined: Thu Nov 01, 2012 12:48 pm

Post by 2468 » Thu Nov 08, 2012 7:48 pm

Hey,
Can someone please confirm my answers before I submit it....correct me if i am wrong instead of beating around the bush! :D

explantion 1= lane 2
2= wrong interpretation
3= lane 3
4= lane 1
5= lane 6
6= wrong interpretation
7= lane 3


Which of the following protocols modifications could remove the high molecular weight bands?
1. Increase the EDTA concentration

2. Expose the cells to the detergent for longer time

3. Increase the concentration of the nuclease

4. Decrease the concentration of the nuclease

5. Increase the incubation time with the nuclease.

6. Increase the number of cells used

I've put down 1,2,3,5

Thank u!

dinnuparus
Garter
Garter
Posts: 1
Joined: Sun Feb 05, 2017 3:34 am

Re: Chromatin experiment

Post by dinnuparus » Sun Feb 05, 2017 3:40 am

do on more cells may be effect some infection controll the infection occurring time. Androdumpper APK finally said Androdumpper.com/ Cinema Box for iPad

Post Reply

Who is online

Users browsing this forum: No registered users and 5 guests