T&A™ Cloning Kit
Description

Molecular cloning assisted by vectors is the most popular and common method to obtain genes of interest. Gentaur 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
  • Vector can be supplied in dried form (# FLC002-20R)
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 × 108 cfu/μg DNA
  • The colony number ratio of self-ligation control to positive control is less than 15%
  • The colony number of positive control is more than 500 when the transformation efficiency of competent cells is 5 × 108 cfu/μg DNA
  • The ligation correctness with the control insert into T&A™ Cloning Vector is more than 87.5%

 

 

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 439
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 incorporate 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.
Figure 1. Map and Sequence reference points of the
T&A™ Cloning Vector

Figure 2. Cloning site of T&A™ Cloning Vector

Figure 3. Restriction enzyme sites of T&A™ Cloning Vector

 

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)

 

Store at

 

# FLC002-20P (20 preps)
T&A Cloning Vector(Dried) 4 tubes*
(*Dissolves with 10 μl/tube of ddH2O prior to use)

 

Store at

 

# FYC004-20P (20 preps)
# FYC001-20P 1 set
# FYE707-50-20VL 1 set
# FYC005-20P (20 preps)
# FYC001-20P 1 set
# FYE607-10VL 2 set
# FYC006-20P (20 preps)
# FYC001-20P 1 set
# FYE607-10VL 2 set
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