Navigation Links
InstaGene Matrix: Prepare DNA Templates for PCR With No Phenol/Chloroform Extractions and No Deproteinization Steps, Rev C


Prepare DNA from blood, cultured cells, or bacteria:
Just incubate sample with InstaGene matrix at 56C for 1530 min, then boil for 8 min.

InstaGene matrix absorbs cell lysis products, producing an improved substrate for PCR amplification.

DNA Preparation From Whole Blood
Use whole blood samples that are frozen, refrigerated, or collected fresh.
1. Add 36 l of whole blood to 1 ml of autoclaved double distilled water in a 1.5 ml microfuge tube. Mix by inverting the tube several times.

2. Incubate the tube at room temperature for 1530 min.

3. Spin at 10,00012,000 rpm for 23 min.

4. Carefully remove all but 2030 l of the supernatant. Do not disturb the pellet.

5. Add 200 l of InstaGene matrix to the pellet and incubate at 56C for 1530 min.

Note: InstaGene matrix should be mixed at moderate speed on a magnetic stirrer to maintain the matrix in suspension. The pipet tip used should have a large bore, such as on a 1,000 l pipet tip (Bio-Rads TBR tips, catalog #223-9350 and 223-9351).

6. Vortex at high speed for 10 sec. Place the tube in a 100C heating block or boiling water bath for 8 min.

7. Vortex at high speed for 10 sec. Spin at 10,00012,000 rpm for 23 min.

8. Use 20 l of the resulting supernatant per 50 l PCR reaction. Store the remainder at -20C. Repeat step 7 when reusing the InstaGene DNA preparation.

Note: It is important to store the prepared sample at -20C when not in use.

DNA Preparation From Cultured Mammalian Cells
1. Pellet 200 l of cells suspension from media in a microfuge tube. Spin at 10,00012,000 rpm for 1 min and remove the supernatant.

2. Resuspend cells in 1 ml of 1x PBS and centrifuge for 1 min at 10,00012,000 rpm.

3. Resuspend cells in autoclaved water at 2030 cells/l.

4. Add 20 l of this cell suspension to 200 l of InstaGene matrix. Incubate at 56C for 1530 min.

Note: InstaGene matrix should be mixed at moderate speed on a magnetic stirrer to maintain the matrix in suspension. The pipet tip used should have a large bore, such as on a 1,000 l pipet tip.

5. Follow the procedure for DNA preparation from whole blood, steps 68.

DNA Preparation From Bacteria
The protocol described below is for the preparation of genomic DNA or episomal DNA from bacteria.
1. Pick an isolated bacterial colony and resuspend it in 1 ml of autoclaved water in a microfuge tube.

2. Centrifuge for 1 min at 10,00012,000 rpm. Remove the supernatant.

3. Follow the DNA preparation procedure for whole blood, steps 58.


back to top
'"/>

Source:


Page: All 1 2

Related biology technology :

1. Preparation of PCR Quality Template DNA from a Wide Range of Tissues Using InstaGene Matrix
2. Prepare siRNA and miRNA Probes in Just 1 Hour
3. Rapidly Prepare Probes and Markers for Small RNA Analysis
4. TripleMaster and Perfect gDNA Blood Mini Kit Team Up to Amplify Long gDNA Templates
5. Using the TripleMaster PCR System for Robust Amplification of GC-Rich DNA Templates
6. Inactivation of Contaminating Templates in PCR Using High Intensity Ultraviolet Light
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:10/12/2017)... ... 12, 2017 , ... BioMedGPS announces expanded coverage of SmartTRAK ... module, US Hemostats & Sealants. , SmartTRAK’s US Market for Hemostats and Sealants ... sealants and biologic sealants used in surgical applications. BioMedGPS estimates the market will ...
(Date:10/11/2017)... Charlotte, N.C. (PRWEB) , ... October 11, 2017 ... ... ARCS® Foundation President Andi Purple announced Dr. Suneel I. Sheikh, the ... Laboratories ( ASTER Labs ), Inc. has been selected for membership in ...
(Date:10/11/2017)... ... , ... ComplianceOnline’s Medical Device Summit is back for its 4th year. The ... Francisco, CA. The Summit brings together current and former FDA office bearers, regulators, industry ... officials from around the world to address key issues in device compliance, quality and ...
(Date:10/11/2017)... Netherlands and LAGUNA HILLS, Calif. ... Institute of Cancer Research, London (ICR) ... MMprofiler™ with SKY92, SkylineDx,s prognostic tool to risk-stratify patients with ... known as MUK nine . The University of ... which is partly funded by Myeloma UK, and ICR will ...
Breaking Biology Technology:
(Date:7/20/2017)... July 20, 2017 Delta (NYSE: DAL ) customers ... Delta aircraft at Reagan Washington National Airport (DCA). ... Delta launches biometrics to board aircraft ... Delta,s biometric boarding pass ... is now integrated into the boarding process to allow eligible Delta ...
(Date:5/16/2017)...  Veratad Technologies, LLC ( www.veratad.com ), an innovative ... verification solutions, announced today they will participate as a ... thru May 17, 2017, in Washington D.C.,s ... Identity impacts the lives of billions ... evolving digital world, defining identity is critical to nearly ...
(Date:4/18/2017)...  Socionext Inc., a global expert in SoC-based imaging and computing ... M820, which features the company,s hybrid codec technology. A demonstration utilizing ... Inc., will be showcased during the upcoming Medtec Japan at Tokyo ... Las Vegas Convention Center April 24-27. ... Click here for an image ...
Breaking Biology News(10 mins):