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:8/16/2017)... and OXFORD, England , Aug. 16, ... consortium for biotech executive search and leadership development, and Virdis ... sectors, have created an exclusive alliance that enables clients to ... "For our clients here in the ... unparalleled access to a diverse population of leadership talent throughout ...
(Date:8/15/2017)... Boston, MA (PRWEB) , ... August 15, 2017 , ... ... unmet need that has compromised these disciplines for more than half a century. ... cannot be counted. It is widely known that molecular tags developed for this ...
(Date:8/14/2017)... (PRWEB) , ... August 14, 2017 , ... ... characterized and performing antibodies. Key researchers in the antibody community have recently come ... and consistency for antibodies in the laboratory. , The team at ...
(Date:8/11/2017)... ... ... Algenist continues to disrupt the skincare industry with today’s debut of GENIUS ... is the key structural element skin needs to maintain its youthful appearance and Algenist ... First to market with proprietary collagen water active , ...
Breaking Biology Technology:
(Date:7/20/2017)... , July 20, 2017 Delta (NYSE: DAL ... board any Delta aircraft at Reagan Washington National Airport (DCA). ... Delta launches biometrics to ... Delta,s biometric ... Sky Club is now integrated into the boarding process to allow ...
(Date:5/23/2017)... robotic gym for the rehabilitation and functional motor sense evaluation of lower ... . The first 30 robots will be available from June in ... The technology was developed and patented at the IIT laboratories and has ... to a 10 million euro investment from entrepreneur Sergio Dompè. ... ...
(Date:4/24/2017)... -- Janice Kephart , former 9/11 Commission ... LLP (IdSP) , today issues the following statement: ... 6, 2017 Executive Order: Protecting the Nation ... instilled with greater confidence, enabling the reactivation of ... are suspended by until at least July 2017). ...
Breaking Biology News(10 mins):