Dear all,
.in my lab i need to isolate pet28a and i use Alkaline Lysis protocol ..but i cannot isolote this plasmid via alkaline lysis when i finish this method and run electrophoresis gel i cannot see any band in gel ..i don't know Why can not I isolate this? :(
and yesterday when i isolate pet 28a and run I saw a weak band and when i use Restriction enzyme to digest this The band disappeared ..and i do not know Why is this happening ؟
best regards
If you want to solve the problem as quick as possible get ahold of an experienced colleague and request her/his help. Remote troubleshooting is too hard with the scant information you are providing, and the list of things that might have gone wrong is long, starting from faulty antibiotic selection during culture and ending in poor EtBr staining during gel electrophoresis.
Is this happening only with this plasmid, or are you unable to isolate other plasmids as well?
See this link may be helpful
http://www.clontech.com/xxclt_ibcGetAttachment.jsp?cItemId=17520
Greeting
- Badges
- Science topic
- Similar topics
- Biological Science
- Kinesiology
- Running