20 μg of total RNA was analyzed on a 15% denaturing polyacrylamide-urea gel. miR-100 and HAGLROS were analysed on a 1.2% agarose gel in 1 × MOPS solution containing 1% formaldehyde. RNAs were separated by electrophoresis and transferred onto a Hyoid-N+ nylon membrane (GE healthcare, Shanghai, China). The membranes were incubated with hydration buffer containing probes in the labelling reaction system (20 mL) containing 2 mL of 10 × T4 PNK ligase buffer and 1 mL of T4 poly nucleotide kinase (NEB, Ipswich, MA, USA) and 2.5 mL of γ-[32 P]-ATP. Then the membranes were pre-hybridized in the hybridization buffer at 65 °C for 1 h and hybridized overnight at 65 °C. The membranes were then exposed to the phosphor screen for at least 12 h to show the bands.
T4 pnk ligase buffer
Manufactured by New England Biolabs
Sourced in United States
T4 PNK ligase buffer is a solution used in molecular biology applications to provide the optimal conditions for the enzymatic ligation of DNA fragments. It contains the necessary cofactors and reagents to facilitate the activity of T4 polynucleotide kinase (T4 PNK) and T4 DNA ligase enzymes.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using t4 pnk ligase buffer
1
Northern Blot Analysis of miR-100 and HAGLROS
2
Northern Blot Analysis of miRNA, mRNA, and lncRNA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For all miRNA samples, 20 μg of total RNA was analysed on a 17% denaturing polyacrylamide‐urea gel. mRNAs and lncRNAs were analysed on a 1.2% agarose gel in 1× MOPS solution containing 1% formaldehyde. RNAs were separated by electrophoresis and transferred onto a Hyoid‐N+ nylon membrane (Habersham, Freiburg, Germany). The membranes were incubated with hydration buffer containing probes in the labelling reaction system (20 mL) containing 2 mL 10× T4 PNK ligase buffer and 1 mL T4 poly nucleotide kinase (NEB, Ipswich, MA) and 2.5 mL γ‐[32P]‐ATP. Membranes were pre‐hybridized in the hybridization buffer at 65°C for 1 hour and hybridized overnight at 65°C. The membranes were then exposed to the phosphor imager screen for at least 12 hours.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!