SampleIN™ Direct PCR Kit

Applications


  • Fast direct PCR without template purification

  • Mouse genotyping and knockout analysis

  • Direct PCR from mouse tail or ear, mammalian tissues (including FFPE)

  • Direct PCR from hair follicle, buccal swabs, plant samples

  • Direct Blood PCR (including EDTA or FTA samples)

Benefits


  • Ready-to load PCR in 50 minutes without template purification

  • Single-tube 15 min DNA extraction combined with fast hot-start PCR

  • Red dye in the PCR master mix for direct gel loading

  • High yields under standard or fast cycling conditions

  • Success with GC/AT rich templates

Catalog. # Product Name Size Price Qty
DPK0101 SampleIN™ Direct PCR Kit 80 r of 50 µl
€98.00
DPK0105 SampleIN™ Direct PCR Kit 400 r of 50 µl
€399.00

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Description

SampleIN™ Direct PCR Kit is a premium tool for a fast direct PCR eliminating the need of tedious template purification. The kit is excellent for direct PCR from mouse tail or ear, mammalian tissues, hair follicle, buccal swabs and blood.

Rapid 15 min DNA extraction using DPK Lysis and Protease Buffers in a single tube generates PCR template extract which is further amplified under fast cycling conditions with a hot-start Taq master mix that includes red dye for direct gel loading.

In a 2% agarose TAE gel the red dye migrates with~350 bp DNA, in 1% agarose TAE gel with ~ 600 bp DNA fragments.

The ALLin™ HS Red Taq Mastermix includes a hot start Taq DNA Polymerase what ensures high yield, specific, low background amplification. Mix components allow for a fast PCR cycling and increase success when working with complex templates or multiplexing. Generated A-tailed PCR products are suitable for ligating into TA cloning vectors, sequencing and other applications.

 

 highQu Customer experience with the SampleIN Direct PCR Kit

Protocols

Print version of the protocol: Product Insert SampleIN™ Direct PCR Kit
Sample Guidelines

 

Sample amounts can be slightly increased for better yields, but too much material may cause inefficient lysis and PCR inhibition.

Sample (fresh or frozen) Amount Extraction vol.
Mouse tail 2 mm or 3-5 mg 100 µl
Mouse ear 2 mm2 or 3-5 mg 100 µl
Mammalian tissue 5 mg 100 µl
FFPE Tissue 2 mm2 of 10 µm section 100 µl
Blood (fresh/EDTA) 2 µl 100 µl
Blood Guthrie cards 2 mm2 100 µl
Blood FTA/FTA Elute cards 2 mm2 100 µl
Hair follicle 2 follicles 100 µl
Buccal swab 1 swab 300 µl
 
Sample extraction procedure:
  • Take typical measures to prevent contamination, keep your bench clean, wear gloves, and use sterile tubes.
  • Thaw DPK Buffers at room temperature. Mix well before use.
  • Prepare a 100 µl extraction reaction in a sterile vial (use 3x larger volumes of all reagents for buccal swab):

 

Sample As indicated above in SAMPLE GUDELINES
DPK Lysis Buffer, 5X 20 μl
DPK Protease Buffer, 10X 10 μl
PCR Water (not supplied) 70 μl
  • Mix very gently. Place into the thermal block/water bath set like:
Lysis 75°C - 5 min. Vortex twice during lysis.
Protease inactivation 95°C - 10 min

 

  • Add 900 µl of PCR Water. Centrifuge 1 min to pellet cell debris.
  • Remove supernatant into the sterile tube.
  • Store it at -20°C for several months or use immediately for PCR.
 
Perform PCR reaction:

 

  • Include a no-template control and positive control in parallel.
  • Thaw and keep PCR reagents on ice. Mix well before use.

 

Rev. & For. Primers 0.1-0.4 µM final each (≤ 2 µl of 10 µM)
Template 1-5 µl of extraction supernatant
PCR Water to 25 μl
ALLin™ HS Red Taq Mastermix, 2X (also available separately)
25 µl

Mix gently. Place into the PCR instrument set like:

Initial denaturation 1 cycle: 95°C - 2 min
Denaturation 40 cycles: 95°C - 15 sec
Annealing 40 cycles: 55-65°C – 15 sec
Extension 40 cycles: 72°C – 15 sec/kb
(90 sec for multiplex)

 

  • In a 2% agarose TAE gel the red dye migrates with~350 bp DNA, in 1% agarose TAE gel with ~ 600 bp DNA fragments.
  • Store probes for short time on ice, for long at -20°C.

Specifications

Cat.

Size

Components

Composition

DPK0101 80 r of
50 µl
1.6 ml - DPK Lysis Buffer, 5X0.8 ml - DPK Protease Buffer, 10X
2x 1 ml - ALLin™ HS Red Taq Mastermix, 2X
DPK Lysis Buffer, 5X contains all components required for an efficient lysis of mammalian tissue samples.DPK Protease Buffer, 10X contains proteases to eliminate sample proteins.1X ALLin™ HS Red Taq Mastermix contains hot-start enzyme, 0.25 mM dNTPs, 3 mM MgCl2, enhancers, stabilizers, red electrophoresis tracking dye and density reagents for gel loading
DPK0105 400 r of
50 µl
5x 1.6 ml - DPK Lysis Buffer, 5X5x 0.8 ml - DPK Protease Buffer, 10X
10x 1 ml - ALLin™ HS Red Taq Mastermix, 2X


Storage


  In the dark at -20°C.

Resources

 

Product Insert