Help on PCR purification - (May/02/2006 )
Help. I run my pcr products tru Qiagen column for purification without doing any spectro reading to it assuming that it would be better to do after purification. Now I'm having problem reading spectro for concentration before printing on slides for my microarray. The reading was way off even to zero(concentration). I had it check using electro and the bands were still there. Anybody had this problem before? Then I check some of my pcr product(non purified), and concentration was ok( around 200-400ug/ml). Should I just print it and check for donut problem?
If the quality of DNA looks good on the gel and you can estimate the concentration you should be fine. You can use a mass ladder to estimate concentration
Are you using a plate reader or cuvette? If reading on a plastic multiwell plate, make sure your plate is UV transparent. Always check for bubbles.
Are you using a plate reader or cuvette? If reading on a plastic multiwell plate, make sure your plate is UV transparent. Always check for bubbles.
I use plate reader and it came up with error for 280 and 320nm. Then i use cuvette and the reading was very low even to zero concentration. Yup I checked and always with no bubbles. Thanks
