An optimized recipe for cloning of the polymerase chain reaction-amplified DNA inserts into plasmid vectors
Küçük Resim Yok
Tarih
2000
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Acta Biochimica Polonica
Erişim Hakkı
info:eu-repo/semantics/closedAccess
Özet
This study compares a number of parameters that are important in the ligation of the polymerase chain reaction-amplified DNA inserts into plasmid vectors and their efficient transformation to bacterial cells. The parameters covered were: T4 polynucleotide kinase treatment followed by either the large fragment of E. coli DNA polymerase or T4 DNA polymerase reactions, the amount of T4 DNA ligase, temperature and duration of ligation, molar ratio of insert to vector as well as the total DNA concentration. The results show that the T4 polynucleotide kinase-treated group without further enzymatic manipulation, at an insert to vector ratio of 3:1 gave the highest recombination efficiency when 10 mu g/ml DNA and 20 units T4 DNA ligase were applied for ligation for 12 h at 4 degrees C.
Açıklama
Anahtar Kelimeler
cloning, PCR, bacterial transformation
Kaynak
Acta Biochimica Polonica
WoS Q Değeri
N/A
Scopus Q Değeri
N/A
Cilt
47
Sayı
3
