ࡱ> tg=` Sbjbj (((<8DD</2PLp/r/r/r/r/r/r/$#1h3f/(`@@//R(p/p/,h"(-D Itn,./0/-33 -3(-(//^/<<<D <<<<<< Using Saliva Sponges to Collect DNA Samples from Infants & Young Children When using sponges to collect DNA please use the following instructions and disregard the instructions in the container packaging. Introduction Saliva can be collected from most adult donors and children by following the user instructions for spitting into the OrageneTM DNA Self-Collection Kit. However, infants and young children are unable to spit the required 1-2 mL of saliva. DNA Genotek has developed a collection procedure for infants and young children that uses saliva sponges and the Oragene DNA Self-Collection kit to collect a sample. DNA yield is proportional to saliva volume. The saliva sponges included in the kit are ideal for collecting the relatively large amount of saliva present in the cheek pouches of young children. More DNA can be collected by taking several saliva sponge samples from each child over a period of time. Preparing for Saliva Collection Caution should always be used when inserting anything into a childs mouth. Do not leave the child unattended when using the sponges. We recommend that you only use the sponges provided and that you do not substitute with other sponges or swabs. Although not required, the child may drink water or brush teeth with water before the collection. After rinsing or brushing, wait 10 minutes before collecting a saliva sample. If the child is nursing, wait 15-20 min after feeding before collecting a saliva sample. Some children find that the saliva sponge tickles their gums. Depending on the age of the child, it may be helpful to explain ahead of time that the collection will be a fun experience that will not take long and that the sponge may tickle. Try not to rub directly on the childs teeth to minimize the amount of bacteria transferred to the sponge. If a donor can provide some saliva, but not the full amount, through spitting, it is perfectly acceptable to combine saliva from spitting together with saliva sponges in the same Oragene kit. If the donor is unable to provide sufficient saliva within the recommended 10-15 min, the following procedure can be tried. Securely cap the Oragene vial after 15 min this will release the Oragene fluid. Mix the contents of the vial gently by inversion 5 times. When a second collection is to be attempted (this can be days later if convenient), place the vial on a flat surface and carefully open the vial, taking care to avoid spilling any of the liquid. Place the cap on a flat surface with the inside facing upwards. Proceed with further collection of saliva using the remaining sponges. Extra care should be taken when cutting off the sponge from the handle to avoid spillage. A maximum of 5 sponges should be used per Oragene vial. Collecting Saliva  EMBED PBrush Place the saliva sponge into the childs mouth in the cheek pouch (the space between the gums and the inner cheek). Gently move the saliva sponge around the upper and lower cheek pouches on both sides of the mouth to soak up as much saliva as possible. There is no need to scrape the inner cheek with saliva sponges simply collect as much saliva as possible from the cheek pouches. The sponge will absorb more saliva if it is left in the childs mouth for a longer time (up to 60 seconds).  EMBED PBrush Once collected, cut the sponge into the blue base of the Oragene kit as follows. Place the sponge firmly against the bottom of the kit between the tooth and the kit wall (see picture below). This action will ensure that the sponge tip remains in the container during the cutting action. Using the scissors provided, cut the narrow part of the handle just above the sponge. Recycle/discard the plastic handle. If only one saliva sponge sample is to be collected, proceed to step 4.  For the collection of up to 5 saliva sponge samples from the same child, repeat steps 1 and 2. Follow the sequence shown in the diagram below. A rest period of about 5 min between each collection of 2 sponges is helpful. To prevent the saliva samples from drying out, cap the vial (see step 4) within 15 min of the first collection. If you have not had a chance to collect all 5 sponges within 15 minutes, you may carefully re-open the kit. If you remove the cap be sure that the inside is facing upwards when putting it on any surface. Do not spill the contents. Follow these steps for collecting multiple sponges:  EMBED PBrush  Carefully cap the kit and tighten it firmly. Once the Oragene liquid is released from the cap, it will preserve the DNA collected by the sponge(s).  EMBED PBrush   Invert gently 5 times to mix the sample. 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