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260/280 and 260/230 readings...


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6 replies to this topic

#1 scoob00

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Posted 07 September 2009 - 02:54 AM

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 :blink:

L

#2 Clare

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Posted 08 September 2009 - 01:02 AM

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



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



#3 jiajia1987

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Posted 08 September 2009 - 01:16 AM

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



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.

#4 scoob00

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Posted 08 September 2009 - 07:44 AM

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

#5 jiajia1987

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Posted 08 September 2009 - 05:00 PM

just for your information, i use qiagen kits.

#6 scoob00

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Posted 14 September 2009 - 01:59 AM

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

Thanks again!!

:)

#7 Clare

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Posted 22 September 2009 - 05:41 AM

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

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

Thanks again!!

:rolleyes:






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