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Molecular Biology

Extract-N-Amp™ Tissue PCR Kit FAQs

Can the isolated DNA be used in applications other than PCR such as restriction digestion or Southern blots?
Can the PCR product amplified with the Extract-N-Amp Tissue PCR Kit be used for other downstream applications such as sequencing?
How long can the extracted or neutralized DNA be stored?
What is the largest or smallest DNA that can be extracted and amplified using this kit?
Can RNA from the samples be used in RT-PCR?
Will this kit work with real-time quantitative PCR?
If the tissue or mouse tail does not completely digest, will the extract work?
Does the user need to provide any reagents to use this kit?
Is it possible to extract the same mouse-tail piece twice?
I would like to perform more amplifications than the kit can support. Can I buy the PCR mix separately?
Can I optimize my PCR conditions?

Can the isolated DNA be used in applications other than PCR – such as restriction digestion or Southern blots?
The Extract-N-Amp and REDExtract-N-Amp Tissue PCR Kits are a balanced system of extraction and amplification designed especially for PCR. The DNA prepared with Extract-N-Amp™ may not be optimal for other downstream applications.

Can the PCR product amplified with the Extract-N-Amp Tissue PCR Kit be used for other downstream applications – such as sequencing?
Yes. We have obtained good sequencing data from PCR products generated with the Extract-N-Amp Tissue PCR Kit after the PCR product has been cleaned up using the GenElute™ PCR Clean-up Kit (Product No. NA1020).

How long can the extracted or neutralized DNA be stored?
The extracts are stable at 4 °C for at least 6 months. Mouse-tail extracts are stable at 37 °C for 10 weeks. This accelerated study would indicate that they would be stable for approximately 2 years at 4 °C.

What is the largest or smallest DNA that can be extracted and amplified using this kit?
We have amplified PCR products as small as 125 bp and as large as 5 kb.

Can RNA from the samples be used in RT-PCR?
No. RNA is degraded during the tissue extraction procedure.

Will this kit work with real-time quantitative PCR?
Absolutely! The amplification plots and the data demonstrating extract stability were obtained by real-time quantitative PCR.

If the tissue or mouse tail does not completely digest, will the extract work?
Absolutely! The Extract-N-Amp Tissue PCR Kit was developed to extract ample DNA for PCR from the tissue or mouse tail in a quick 15-minute procedure. It is not expected for the tissue or mouse tail to be completely digested in 15 minutes.

Does the user need to provide any reagents to use this kit?
No. All reagents for extraction and PCR are included in the Extract-N-Amp Tissue PCR Kit. All the user needs to provide is primers for the PCR reaction.

Is it possible to extract the same mouse-tail piece twice?
After extracting DNA from a mouse tail using the Extract-N-Amp Tissue PCR Kit, the tail piece can be removed and extracted again using the same procedure. The results from the second extraction are comparable to the first.

I would like to perform more amplifications than the kit can support. Can I buy the PCR mix separately?
Yes. The 2x PCR mixes, both REDExtract-N-Amp PCR Readymix (Product No. R4775) and Extract-N-Amp PCR Readymix (Product No. E3004), can be purchased separately.

Can I optimize my PCR conditions?
Yes. You could adjust your PCR reaction knowing that the available Mg2+ in the PCR mix is 2.5 mM, but this may interfere with the optimal performance of the kit.