KG205-Mouse Tissue Direct PCR Kit

Mouse Tissue Direct PCR Kit
——Fastest DNA Extraction from Mouse Tissue Followed by PCR

Mouse Tissue Direct PCR Kit
Cat. No. 4992531/4992532

Storage
Tissue Lysis Buffer and Digestive Enzyme can be stored for 12 months at
room temperature (15-30°C); 2× Dir PCR MasterMix can be stored for up
to 12 months at -30~-15°C, and repeated freeze-thawing will not affect its
activity.

Introduction
The kit adopts a unique packaging system, which contains all reagents
for rapidly extraction of mouse tissue genome DNA and subsequent
PCR amplification. It is suitable for extracting genome DNA from mouse
tail, ears, toes and other tissues in one step and for subsequent PCR
amplification and detection. The whole extraction process does not include
homogenization, smashing, overnight digestion, phenol/chloroform
extraction, DNA precipitation or column purification, etc. Stable and
reliable results can be achieved with simple and fast experimental
operation.
The 2× Dir PCR MasterMix provided by the kit is a PCR reagent with
high amplification compatibility and can perform high-efficiency and
specific amplification without completely removing impurities such as
proteins. The premixed MasterMix comprises antibody modified Taq
DNA polymerase, dNTPs, MgCl2
, reaction buffer, PCR reaction enhancer
and stabilizer. Only a crude extraction template and primers are needed
for subsequent PCR detection. The premixed MasterMix has the
characteristics of simple operation, high sensitivity, strong specificity, good
stability, etc., and is especially suitable for high-throughput detection and
screening. The MasterMix contains electrophoretic dye, which can be used
for electrophoresis detection directly after the reaction. The PCR product
has a dA-tailing, which can be used for TA cloning.

Features
Easy and fast: Suitable for one-step gene identification from mouse tail,
ears, toes and other tissues.
High specificity:The Taqpolymerase used in this product is antibody
modified HotStart polymerase, with high amplification specificity and high
affinity for template and primers, and is especially suitable for genotyping
and transgene identification.
Gene detection: This product is easy to operate and produces reliable
results, which is especially suitable for high-throughput analysis and
detection of mice.

Important Notes Before Starting

1. Avoid repeated freezing and thawing of the sample, otherwise the
   extracted DNA fragments will be smaller and the extraction yield will be
   decreased.
2. The Tissue Lysis Buffer should be stored at room temperature. If a
   precipitate has formed in Buffer, please place the buffer at room
   temperature or warm at 37°C for 10 min to dissolve the precipitate.
3. The 2× Dir PCR MasterMix provided by this product is 2× Mix. Template,
   primers and sterilized water should be added to make up to a 1×
   solution before use.

Protocol

• Before using this kit, please carefully check whether there is precipitation
  in Tissue Lysis Buffer. If precipitation forms, please fully balance the
  buffer at room temperature until the precipitation is completely
  dissolved, or redissolve the precipitate in 37°C water bath and shake
  well before use. Dissolve the Tissue Lysis Buffer and store at room
  temperature.
• Prepare digestion buffer according to the following formula:
  Note: Please prepare the digestion buffer right before use to ensure
  the activity of Digestive Enzyme.

• Take a small amount of mouse tissue samples (about 5~10 mg) into a
  1.5 ml centrifuge tube and add 100 µl of digestion buffer. Ensure that
  the tissue samples are completely soaked in the digestion buffer and
  incubate at 65°C for 30 min. During this period, flick the tube bottom
  every 10 min to improve digestion efficiency.
• After digestion is completed, instantaneously centrifuge the mixture and
  process at 95~100°C for 5 min.
• Take 1 µl supernatant for PCR reaction. Please refer to the PCR system
  and amplification procedure below:
  

PCR Product Detection
After the reaction is completed, take 5-10 μl of the reaction product for
agarose gel electrophoresis detection.
Note: Examples are provided for reference only. The actual reaction
conditions vary according to the structure of templates, primers, etc. The
best reaction conditions should be set according to the actual situation.
If liquid is found on the tube wall or cover during operation, centrifuge
instantaneously to the bottom of the tube.

KG205-Mouse Tissue Direct PCR Kit

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This product is for scientific research use only. Do not use in medicine, clinical treatment, food or cosmetics.

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