In general, when the water production rate or desalination rate of the RO system is reduced to 10-15%, it is necessary to perform CIP treatment on the RO membrane module. The following CIP steps are for your reference: 1. Water Cleaning
Full Answer
What makes a CIP system effective?
The effectiveness of a CIP system first relies on four factors: mechanical action, chemical action, temperature, time for cleaning-in-place. These four factors must be considered altogether: any reduction in one of the quadrants in cleaning in place must be balanced by increasing one or more of the other factors.
What is cleaning in place (CIP)?
The Cleaning In Place (CIP) process is defined as the process of cleaning the interior of machines without removing or dismantling parts of the equipment.
What are the CIP steps for the RO system?
The following CIP steps are for your reference: 1. Water Cleaning Rinse RO membrane modules and system pipes with water at a temperature of 60-80℃ to remove dirt and other residual substances on the surface.
What happens if you use too much CIP?
If residual CIP activity carries through to the ligation, it will dephosphorylate the insert, preventing ligation from occurring. 4. Anecdotally, overdoing the alkaline phosphatase treatment can damage the DNA.
What does CIP treatment do?
CIP is commonly used to remove 5'-phosphate groups from DNA, RNA and both ribo and deoxy-ribonucleoside triphosphates. Removal of 5'-phosphates is very useful in preventing self-ligation of cleaved DNA vectors. This property greatly reduces background (plasmids without insert) in cloning procedures.
What is the ideal condition for CIP enzymatic activity?
CIP activity is optimal in NEBuffers 2, 3 or 4 and should not be use in NEBuffer 1 (50% activity). Slightly lower activity may be observed in any reaction buffer containing a total salt concentration of less than 50 mM, under these conditions a 2-fold excess of CIP is recommended.
Why is alkaline phosphatase treatment necessary in cloning?
Alkaline phosphatases are routinely used to reduce the background from empty, religated vectors during cloning of DNA fragments, since dephosphorylated DNA termini cannot be ligated by DNA ligase. The phosphatase treatment will effectively reduce the background of “empty” clones by >95%.
What is the point of using calf intestine phosphatase?
Calf intestinal alkaline phosphatase (CIAP) catalyzes the hydrolysis of 5´-phosphate groups from DNA, RNA, and ribo- and deoxyribonucleoside triphosphates. This enzyme is used to prevent recircularization and religation of linearized cloning vehicle DNA by removing phosphate groups from both 5´-termini (1–5).
How does CIP prevent self ligation?
CIP works by removing the phosphate group from the 5′ end of linearised DNA, which means you can use it to dephosphorylate your vector molecule to prevent it from self-ligating and giving you the headache of a high empty vector background after ligation and transformation.
Does CIP work in conventional restriction enzyme buffers?
-does CIP work in conventional restriction enzyme buffers? (to answer the previous 3 questions: you may find relevant information online!) CIP is active in all restriction enzyme NEBuffers 1, 2, 3 as well as NEBuffers 1, 2, 3, 4, NEBuffer for EcoRI, and CutSmart Buffer. It can be added directly to digested DNA.
Why do I need to dephosphorylate my vector?
Dephosphorylation is only necessary for the vector backbone. You are simply trying to prevent the backbone closing on itself and giving you colonies that can propagate this empty vector (absent of your gene of interest).
Is phosphorylation necessary for ligation?
Since the vector has been dephosphorylated, and ligation requires the presence of a 5'-phosphate, the insert must be phosphorylated.
Do I need to dephosphorylate my insert?
For linear vectors with unique 5′ ends, dephosphorylation is not necessary. The dephosphorylation reaction can be performed directly in restriction enzyme buffer so the vector can be cut and dephosphorylated at the same time.
What is CIP enzyme?
Alkaline Phosphatase, Calf Intestinal (CIP) nonspecifically catalyzes the dephosphorylation of 5´ and 3´ ends of DNA and RNA phosphomonoesters. Faster reaction setup (no supplemental additives like zinc required) Flexible reaction conditions (active in any restriction enzyme buffer, no clean-up required)
What is intestinal alkaline phosphatase?
Intestinal alkaline phosphatase (IAP) is secreted by enterocytes and is present on the apical surface. It not only detoxifies bacterial endotoxin lipopolysaccharide (LPS) in the gut lumen and limits intestinal inflammation but also restricts translocation of LPS into systemic circulation.
What is bacterial alkaline phosphatase?
Bacterial alkaline phosphatase (BAP) from Escherichia coli (E. coli) is a key enzyme used in protein/antibody detection and molecular cloning. As it removes terminal phosphate from DNA, RNA and deoxyribonucleoside triphosphates, it is used to lower self-ligated vectors' background.
What is a CIP in RO plant?
How to use CIP in RO Plant? CIP is the abbreviation of Clean In Place, which is a cleaning method for cleaning equipment without changing the position of the equipment. It can not only clean the equipment, but also control microorganisms, so it's widely used in the highly mechanized food and beverage production industries, such as beverage, dairy, ...
What does it mean when a CIP is darker?
If the backwater of the CIP is darker and appears turbid, it means that there are more pollutants and the cleaning fluid must be discharged and reconfigured. 3. Water Cleaning.
What to wear when cleaning a CIP tank?
It is recommended to wear protective glasses, rubber gloves, and protective clothing. Before adding the acid and alkali stock solution to the CIP cleaning tank, the tank must be filled with water in advance.
How long does it take to clean a RO system?
The cleaning time is generally 15-20 minutes.
What is a CIP cleaning device?
CIP cleaning devices are capable of chemically cleaning and overall sanitizing a RO system. The chemicals selected do have to be chosen wisely though. Harsh chemicals can actually cause corrosion necessitating repairs or, worse, the replacement of the overall system. Building a RO system with a CIP component is not standardized though.
Can you delay a CIP cleaning unit?
Those who already are using a RO system should move quickly on having a CIP cleaning component or process installed. You cannot delay having the cleaning unit installed forever. A point of no return can be reached in where the membranes cannot be effectively cleaned.
Can you clean a CIP membrane?
The longer you wait to launch a CIP cleaning, the more trouble the membranes can end up suffering. In particular, cleaning out the membranes becomes much more difficult since there is more to clean….if the membranes can be cleaned at all.
