Protocol

Lysate Preparation:

1.  Collect desired cells (e.g., swab inner cheek, wound, skin, toe nail, etc. with a swab, centrifuge sample, pick bacterial colony, etc.)
   1. 100 ~ 100,000 cells / sample works best
   2. If using cells from an applicator, place cells in ~0.5 mL PBS or water by submerging the applicator in the buffer and twirling the applicator to dislodge cells
2. Add PCRopsis™ 5x Cell Lysis Buffer (cat. no. ******) to make a 1x suspension; mix suspension thoroughly
   1. Example: add 25 μL of 5x Cell Lysis Buffer to a 100 μL cell suspension
3. Heat 0.5 mL cell suspension at 95°C ~100°C for 5~10 minutes to lyse cells; mix suspension thoroughly
   1. TIP: 5 minutes is sufficient for mammalian cells; bacterial and fungal cells may require 10 minutes (or longer) for complete lysis
4. Let lysate cool and directly use lysate with PCRopsis Cell (no need to centrifuge lysate)

Removal of PCR inhibitors from lysate:​

1. Tap PCRopsis Cell tube to ensure the substrate is at the bottom of the tube
2. Add 100~400 μL of lysate to a PCRopsis Cell tube and thoroughly mix lysate with substrate.
   1. TIP: stir sample with pipette tip to ensure all of the substrate is coated by the lysate
   2. TIP: lower volumes of lysate result in samples with the lowest levels of PCR inhibitors
3. Incubate PCRopsis Cell tube with lysate for 10 minutes at room temperature
4. Remove cleaned lysate and use in desired PCR application
   1. TIP: insert pipette tip along the edge of the PCRopsis Cell tube until the very bottom when removing cleaned lysate
   2. TIP: wiggle pipette tip if you notice clogging 
   3. TIP: 1~7 μL of cleaned lysate should be used per 30 μL PCR mixture

PCRopsis Cell is an Alternative to:

1. Invitrogen's ChargeSwitch™ gDNA Normalized Buccal Cell Kit
2. Qiagen's QIAamp® DNA Kit

​Performance

The efficient PCRopsis Cell procedure enables consistent detection of DNA from various cellular lysates. Each lots is carefully tested to ensure maximum quality.

​Principal

PCRopsis Cell is engineered to quickly and simultaneously bind a variety of PCR inhibitors found in cellular lysates. These substrates do not bind to DNA. This approach allows direct detection of DNA from cellular lysates without performing time-consuming DNA isolation protocols, centrifugations or other sample manipulations which may introduce errors, contaminants or skew the representation of DNA fragments.

​Applications

Sequencing / detection of mutations
​Life science research
Livestock breeding
Pedigree genotyping
Veterinary pathogen research
Routine applied testing

Studies with PCRopsis Cell

​(Click on the icons below for more details)

One-Step Clean-Up with
PCRopsis Cell

PCRopsis Cell &
Bacterial Lysate in Urine

PCRopsis Cell + Thermal Lysis vs
​Qiagen's QIAamp® DNA Kit

PCRopsis Cell
with Cell Lysis Buffer +
Thermal Lysis
vs
​​QIAamp® DNA Kit

Human Samples:
PCRopsis Cell
vs
​​QIAamp® DNA Kit

Fungal Samples:
PCRopsis Cell
vs
​​QIAamp® DNA Kit