transgenic mouse genotyping - (Aug/24/2008 )
Hi
Some basic questions.I am doing DNA extraction using Qiagen DNeasy tissue kit,but my DNA concentrations are quite variable such as from 50ng- 250ng/ul.I wish to add just 1 or 2 ul to the pcr reaction mix while doing the PCR,so my question is i)whether variable DNA amounts in different samples affect my result.ii)In genotyping we are just supposed to see the desirable gene product only or quantity of the dna is important,that is should i have equal amounts of DNA in all samples.
-bactin2-
QUOTE (bactin2 @ Aug 25 2008, 10:41 AM)
Hi
Some basic questions.I am doing DNA extraction using Qiagen DNeasy tissue kit,but my DNA concentrations are quite variable such as from 50ng- 250ng/ul.I wish to add just 1 or 2 ul to the pcr reaction mix while doing the PCR,so my question is i)whether variable DNA amounts in different samples affect my result.ii)In genotyping we are just supposed to see the desirable gene product only or quantity of the dna is important,that is should i have equal amounts of DNA in all samples.
Some basic questions.I am doing DNA extraction using Qiagen DNeasy tissue kit,but my DNA concentrations are quite variable such as from 50ng- 250ng/ul.I wish to add just 1 or 2 ul to the pcr reaction mix while doing the PCR,so my question is i)whether variable DNA amounts in different samples affect my result.ii)In genotyping we are just supposed to see the desirable gene product only or quantity of the dna is important,that is should i have equal amounts of DNA in all samples.
Hi,
genotyping is all or none test - any quantity enough for PCR is Ok. May be 50 ng is little for PCR, then just add 2 ul you will be safe.
-MolBio79-
QUOTE (bactin2 @ Aug 25 2008, 10:41 AM)
Hi
Some basic questions.I am doing DNA extraction using Qiagen DNeasy tissue kit,but my DNA concentrations are quite variable such as from 50ng- 250ng/ul.I wish to add just 1 or 2 ul to the pcr reaction mix while doing the PCR,so my question is i)whether variable DNA amounts in different samples affect my result.ii)In genotyping we are just supposed to see the desirable gene product only or quantity of the dna is important,that is should i have equal amounts of DNA in all samples.
Some basic questions.I am doing DNA extraction using Qiagen DNeasy tissue kit,but my DNA concentrations are quite variable such as from 50ng- 250ng/ul.I wish to add just 1 or 2 ul to the pcr reaction mix while doing the PCR,so my question is i)whether variable DNA amounts in different samples affect my result.ii)In genotyping we are just supposed to see the desirable gene product only or quantity of the dna is important,that is should i have equal amounts of DNA in all samples.
Light band - Strong band should not make difference.
But, when there is no band - it might be dubious. Is there really no band or is it too light to be seen?
-Bungalow Boy-