6 replies to this topic

[scoob00]

Hi guys, hope someone can help...

I have just read my colum-cleaned up, excised bands of specific-gene amplified cDNA on a nanodrop. (2 diff samples).

Here are the results;

1: 27.9 ng/ul. 260/280 = 2.06.  260/230 = 0.48.

2: 9.8 ng/ul.  260/280 = 2.10.  260/230 = 0.28.

Obviously the 260/230 readings are horrible. Does this mean my DNA is rubbish? I want to use it for a ligation...

Thanks for any help  

L

[Clare]

I always get rubbish readings after I gel purify my digests (using the QIAGEN kit), but I go and use the DNA anyway and my ligations always work

Clare

scoob00, on Sep 7 2009, 11:54 AM, said:

Hi guys, hope someone can help...

I have just read my colum-cleaned up, excised bands of specific-gene amplified cDNA on a nanodrop. (2 diff samples).

Here are the results;

1: 27.9 ng/ul. 260/280 = 2.06.  260/230 = 0.48.

2: 9.8 ng/ul.  260/280 = 2.10.  260/230 = 0.28.

Obviously the 260/230 readings are horrible. Does this mean my DNA is rubbish? I want to use it for a ligation...

Thanks for any help  

L

[jiajia1987]

Clare, on Sep 8 2009, 05:02 PM, said:

I always get rubbish readings after I gel purify my digests (using the QIAGEN kit), but I go and use the DNA anyway and my ligations always work

Clare

scoob00, on Sep 7 2009, 11:54 AM, said:

Hi guys, hope someone can help...

I have just read my colum-cleaned up, excised bands of specific-gene amplified cDNA on a nanodrop. (2 diff samples).

Here are the results;

1: 27.9 ng/ul. 260/280 = 2.06.  260/230 = 0.48.

2: 9.8 ng/ul.  260/280 = 2.10.  260/230 = 0.28.

Obviously the 260/230 readings are horrible. Does this mean my DNA is rubbish? I want to use it for a ligation...

Thanks for any help  

L

Ditto!

Gel clean up sometimes give me crappy readings. It usually gives worse readings as compared to the PCR clean up kit, but using the cleaned up DNA still works.

[scoob00]

Great, thanks guys!

I'll go ahead with it. I used the Promega Wizard clean up kit. I didn't want to waste my cells etc if the DNA was rubbish to start with - I've already had real problems trying to clone with these two as it is.

Cheers again,
Louise

[jiajia1987]

just for your information, i use qiagen kits.

[scoob00]

Thanks all, just to let you know it all worked perfectly with these readings - juat had 24 positive clones from each line.

Thanks again!!

[Clare]

YAY!!!!!!!!!!!!

scoob00, on Sep 14 2009, 10:59 AM, said:

Thanks all, just to let you know it all worked perfectly with these readings - juat had 24 positive clones from each line.

Thanks again!!