Problems with cDNA - How long can I keep cDNA on 37°C? (Apr/01/2008 )
Hello,
this stupid PCR Machine did not do as I programmed. It kept the whole night on 37°C instead of 60 min. Now my question. Can I still use the cDNA?
I used M-MLV Reverse Transcriptase, RNase H Minus, Point mutation from Promega.
What do you think?
-Sumpf-
QUOTE (Sumpf @ Apr 1 2008, 09:08 AM)
Hello,
this stupid PCR Machine did not do as I programmed. It kept the whole night on 37°C instead of 60 min. Now my question. Can I still use the cDNA?
I used M-MLV Reverse Transcriptase, RNase H Minus, Point mutation from Promega.
What do you think?
this stupid PCR Machine did not do as I programmed. It kept the whole night on 37°C instead of 60 min. Now my question. Can I still use the cDNA?
I used M-MLV Reverse Transcriptase, RNase H Minus, Point mutation from Promega.
What do you think?
Was is machine or programmer error?
At any rate, you'll have to start again. You cDNA will be of very low quality.Ginger
-Ginger Spice-
QUOTE (Ginger Spice @ Apr 1 2008, 10:44 AM)
QUOTE (Sumpf @ Apr 1 2008, 09:08 AM)
Hello,
this stupid PCR Machine did not do as I programmed. It kept the whole night on 37°C instead of 60 min. Now my question. Can I still use the cDNA?
I used M-MLV Reverse Transcriptase, RNase H Minus, Point mutation from Promega.
What do you think?
this stupid PCR Machine did not do as I programmed. It kept the whole night on 37°C instead of 60 min. Now my question. Can I still use the cDNA?
I used M-MLV Reverse Transcriptase, RNase H Minus, Point mutation from Promega.
What do you think?
Was is machine or programmer error?
At any rate, you'll have to start again. You cDNA will be of very low quality.Ginger
*g* no I checked the log file and it was the machine. It stayed on pause but I definitely resumed after I added the Master Mix. This I could see from the log file. But when I came this morning, it was on pause.
-Sumpf-
I tested the cDNA with TaqMan and it worked even better than all my samples before. I got almost perfect triplicates.
-Sumpf-