Gene Cloning: Vectors & Protein Expression

 

T&A™ Cloning Kit

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Multiple Cloning region 434 to 490
LacZ start codon 165
LacZ operator 185 to 201
Lac Z gene 511 to 149
Ori 909-1497
Ampr gene 2528 to 1671
T7 promoter 402 to 419
M13 forward primer 359 to 375
M13 reverse primer 507 to 528
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
  • 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%

Composition

Catalog Number: YSN-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)
Catalog Number: YSN-FYC002-20P (20 preps) 
T&A™ Cloning Vector 40 μl (25 ng/ μl)
Control Insert DNA 10 μl (10 ng/ μl)

DNA Sequence of Multiple Cloning Sites (MCS)

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Ordering Information

T-A Cloning

Catalog Number Product Name
YSN-FYC001-20P T&A Cloning Kit
YSN-FYC002-20P T&A Cloning Vector Kit

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