T&A™ Cloning Kit

Description
Molecular cloning assisted by vectors is the most popular and common method to obtain genes of interest. Yeastern Biotech's T&A™ Cloning Kit offers a quick, reliable and efficient method for cloning a variety of DNA sequences. The T&A™ Cloning Kit contains the T&A™ Cloning Vector and all the reagents needed for ligation. It is a convenient pack for cloning PCR product generated using thermostable DNA polymerases, such as YEAtaq DNA polymerase, which add a single terminal 3'-dA nucleotide overhang. After ligation, the mixture can be used directly for transformation into competent cells (ECOS™) or be purified first to achieve higher transformation efficiency.
Features
  • Fast ligation, completed in only 5 minutes
  • High transformation efficiency
  • More accurate results
  • Accept a wide range of inserts with different sizes
  • Two types of ligation buffers provided for your convenience
  • Allow blue/white screening
  • Contain ampicillin marker for antibiotic selection
  • Include M13 primer sites for convenient sequencing
Map & Sequence Reference Points
 
Multiple Cloning region 434 to 490
LacZ start codon 165
LacZ operator 185 to 201
Lac Z gene 511 to 149
Phage f1 region 2365-2820
Ampr gene 2528 to 1671
T7 promoter 402 to 419
M13 forward primer 359 to 375
M13 reverse primer 507 to 528
β-lactamase coding region 1322 to 2182
Lac operon sequences 151 to 380, 2821 to 2981
* Before the insert incorporates into the T&A™ Cloning Vector, there is only one Hind III site and no Bgl II site. After the incorporation, the T and A nucleotide on the insert will complement the sequence on the vector and generate these two new sites. This merit of T&A™ Cloning Vector makes cloning more economic and convenient.
 
DNA Sequence of Multiple Cloning Sites (MCS)
 
Restriction Enzymes Sites
Restriction Enzymes That Cut The Cloning Vector Once   Restriction Enzymes That DO NOT Cut The Cloning Vector Restriction Enzymes That Cut The Cloning Vector >2 Times
 
Applications
Cloning of terminal 3’-dA nucleotides overhang PCR products up to 5 kb
Quality Control
  • DNA concentration of T&A™ Cloning Vector is 25 ng/μl
  • The absorbance ratio (A260/A280) is between 1.6~2.0
  • The size of T&A™ Cloning Vector is about 2.7 kb
  • The colony number of background control is less than 50 when the transformation efficiency of competent cells is 1 × 108cfu/μg DNA
  • The colony number ratio of self-ligation control to positive control is less than 15%
  • The colony number of the positive control is more than 500 when the transformation efficiency of competent cells is 5 × 108cfu/μg DNA
  • The ligation correctness with the control insert into T&A™ Cloning Vector is more than 87.5%
Specification
Store at

# FYC001-20P (20 preps) 

T&A™ Cloning Vector 40 μl (25 ng/ μl)
Control Insert DNA 10 μl (10 ng/ μl)
yT4 DNA Ligase 20 μl (2U/ μl)
10X Ligation Buffer A 50 μl
10X Ligation Buffer B 50 μl
Forward Primer (M13-F) 50 μl (10 μM)
Reverse Primer (M13-R) 50 μl (10 μM)

# FYC002-20P (20 preps) 

T&A™ Cloning Vector 40 μl (25 ng/ μl)
Control Insert DNA 10 μl (10 ng/ μl)
Manual