Splinkerette pcr is a nice protocol. Haven't tried it myself. I disagree on your assertion that inverse PCR is difficult. I've used it numerous times to identify mouse transgene insertion sites as well as some bacterial transposon integrations. No mechanical shearing needed, only restriction digest. You have to pick the right enzyme(s) depending on your transgene. It has always worked well in my hands.
On Thursday, July 12, 2018 at 9:44:14 AM UTC-7, Skyler Gordon wrote:
-- On Thursday, July 12, 2018 at 9:44:14 AM UTC-7, Skyler Gordon wrote:
Inverse PCR is also going to be difficult unless you have some sort of mechanical DNA shearing method. Common enzymatic and chemical shearing will break the DNA down into pieces that are just too small.I would advise using a splinkerette pcr, or following the protocol that I've attached. Both are simple, rather elegant ways to ensure your PCR products come from the desired location in your genome.-SG
-- You received this message because you are subscribed to the Google Groups DIYbio group. To post to this group, send email to diybio@googlegroups.com. To unsubscribe from this group, send email to diybio+unsubscribe@googlegroups.com. For more options, visit this group at https://groups.google.com/d/forum/diybio?hl=en
Learn more at www.diybio.org
---
You received this message because you are subscribed to the Google Groups "DIYbio" group.
To unsubscribe from this group and stop receiving emails from it, send an email to diybio+unsubscribe@googlegroups.com.
To post to this group, send email to diybio@googlegroups.com.
Visit this group at https://groups.google.com/group/diybio.
To view this discussion on the web visit https://groups.google.com/d/msgid/diybio/5cf21e68-d8a5-465d-b6b9-bbbd77f7445d%40googlegroups.com.
For more options, visit https://groups.google.com/d/optout.
0 comments:
Post a Comment