NEBNext Second Strand Synthesis (dNTP-free) Reaction Buffer

• 产品编号：B6117S      品牌：New England Biolabs       原厂货号：B6117S
• 产品分类：生化试剂 > 生物缓冲液 > 测序缓冲液和溶液
• 应用分类：

描述：

Description:
New England Biolabs supplies a unique 10X reaction buffer for use with NEBNext® Second Strand Synthesis Enzyme Mix when alternative concentrations of deoxynucleotides are required.

Reagents Supplied:
NEBNext® Second Strand Synthesis (dNTP-Free) Reaction Buffer

1X NEBNext® Second Strand Synthesis (dNTP-Free) Reaction Buffer:
20 mM Tris-HCl
12 mM (NH₄)₂SO₄
10 mM MgCl₂
0.16 mM β-NAD
pH 7.5 @ 25°C

 

Quality Control for Current Lot
16-Hour Incubation:
A 50 µl reaction containing this reaction buffer at a 1X concentration and 1 µg of HaeIII digested ΦX174 RF I DNA incubated for 16 hours resulted in no detectable non-specific nuclease degradation.

Endonuclease Activity:
Incubation of this reaction buffer at a 1X concentration with 1 µg ΦX174 RF I DNA for 4 hours at 37°C in 50 µl reactions resulted in less than 5% conversion to RF II.

RNase Activity:
Incubation of this reaction buffer at 1X concentration with 40 ng of a FAM-labeled RNA transcript for 16 hours at 37°C results in no detectable RNase activity as determined by polyacrylamide gel electrophoresis.

Phosphatase Activity:
Incubation of this reaction buffer at a 1X concentration in protein phosphatase assay buffer (1 M diethanolamine @ pH 9.8 and 0.5 mM MgCl₂) containing 2.5 mM p-nitrophenyl phosphate at 37°C for 4 hours yields no detectable p-nitrophenylene anion as determined by spectrophotometric analysis at 405 nm.
 

原厂资料：