EduPrimer™ VNTR DNA Profiling Kit
Contents
1. heterozygous homozygous of alleles w out Alu Total amp alleles ince a Sum 1 00 of um 1 class total alleles Each person has two alleles consequently in calculating allele frequency both copies are taken into account To determine the frequency of the allele with the Alu insert double the number of individuals with homozygous genotypes and add the number of individuals with heterozygous genotypes This sum accounts for all the alleles To determine the frequency of the allele without the Alu insert double the number of individuals with homozygous genotypes and add the number of individuals with heterozygous genotypes This sum accounts for all the alleles Divide this sum Compare with classes around the world Scientists use Alu frequency to study human populations To learn examples and compare the frequencies measured in your class to other classes and populations around the world visit the Allele Server in Cold Spring Harbor s Dolan DNA Learning Center at www bioservers org bioserver EduPrimer DNA Profiling Kit GenoSensor Corp 11 EduPrimer DNA Profiling Kit Background Introduction to PCR In 1983 during his time working at Cetus Corporation Kary Mullis developed a technique that has changed the field of genetics and biological science in general This revolutionary process was termed polymerase chain2. 0 1 Our genome consists of over 3 billion base pairs though and even that small percentage makes a big difference This is important in DNA profiling There are a number of specific regions on our genomes that we can rely on to vary between individuals For this experiment we will be focusing on a Variable Number Tandem Repeat region Throughout our DNA we ve found segments that feature small repeating sequences of DNA The repeating sequence is generally the same between individuals but the number of times it repeats can vary Analyze enough of these segments and you can generate a genetic fingerprint for an individual In the EduPrimer VNTR DNA Profiling kit we focus on the Penta E microsatellite region a VNTR region with a repeating sequence 2 5 base pairs long which features a sequence of AAAGA that can repeat anywhere from 5 to 26 times This means that when a region flanking and including this repeating sequence is amplified by PCR using our primers the region will be between 246 351 base pairs long Each individual will have two bands on the final gel one inherited from each of their parents apart from individuals who might have identical repeating numbered sequences from both parents What this means on the whole is that for a single VNTR site there can be wide degrees of variation and it s unlikely that any two non related persons will share the same band pattern In a real world situation for DNA
3. profiling or fingerprinting many variable regions would be used to be able to accurately distinguish one individual EduPrimer DNA Profiling Kit GenoSensor Corp 6 EduPrimer VNTR DNA Profiling Kit Protocol Preparation 1 2 Set heat block or water bath to 95 C For a heat block it is recommended to add water or sand to ensure proper heat transfer For a water bath be sure tubes are tightly sealed and not fully submerged to avoid contamination Thaw 2x PCR Master Mix on ice Before opening tube spin 10 sec at 6 000 rpm or greater in a microcentrifuge Vortex 10 seconds then spin again for 10 seconds DNA Preparation Do not eat or brush teeth one hour prior to cheek cell collection Wear gloves and handle solutions carefully 1 2 3 11 13 14 EduPrimer DNA Profiling Kit GenoSensor Corp Add 200u l of Solution A red label to a marked microcentrifuge tube Thoroughly roll provided cotton swab inside cheek for no less than 10 seconds to collect cheek cells Place swab into marked tube with Solution A Cut swab above tube to a length that will fit inside the tube Make certain the cap will shut tightly Press the tube against the vortex machine to thoroughly mix the sample for at least 10 seconds Solution A contains components which chemically disrupt cell membranes and begin to unravel proteins Under these conditions the cheek cells will begin to lyse or break open spilling cell conte
4. Alu element Figure 1 The Alu element is a DNA sequence about 300 base pairs long that is repeated one copy at a time almost 500 000 times within the human genome The origin and function of such randomly repeated sequences is not yet known The Alu name comes from the Alu restriction enzyme enzymes that cut DNA at specific sequences recognition site that is found in this sequence The following section reviews the Alu element in more detail PCR Stages The machinery required to perform PCR is known as a thermal cycler The thermal cycler enables the steps of PCR to be automated The reaction involves a repetitive series of cycles each of which consists of template denaturation primer annealing and extension of the annealed primer by Taq DNA polymerase Before beginning DNA amplification genomic DNA is prepared from students cells The students DNA is then added to a mixture of the necessary reagents oligonucleotide primers thermostable DNA polymerase Taq the four deoxynucleotides A T G C and reaction buffer These reagents are pre mixed as a 2X PCR Master Mix in the EduPrimer DNA profiling kit The tubes are placed into the thermal cycler These thermal cyclers contain an aluminum block that holds the samples and can be rapidly heated and cooled across extreme temperature differences The rapid heating and cooling of this thermal block is called temperature cycling or thermal cycling The first step of the PCR temperatur
5. C until the user is ready to proceed to the analysis of the product EduPrimer DNA Profiling Kit GenoSensor Corp 15 Genomic DNA Extraction Genomic DNA Primers Anneal and Polymerase Extents PCR Amplification Agarose Gel Hlectrophoresis Figure 3 Experiment flowchart from start to finish EduPrimer DNA Profiling Kit GenoSensor Corp Troubleshooting Symptom Possible causes Solutions No amplification product Questionable template quality Analyze starting material Inhibitory Substance in reaction Decrease sample volume Insufficient cycle Run additional cycle Incorrect thermal cycler program Verify times and temperatures Errors in heat block incubation Calibrate heating block use sand or water to maximize contact with tube for proper heat transfer Contaminated tubes solutions Autoclave tubes and use filter tips Primer annealing temperature too high Lower annealing temperature in 2 increments Weak bands faint Low concentration of DNA template Increase swabbing time signal thoroughly swab DNA Dye degradation during Light sensitive dyes should be preparation kept in the dark during gel preparation Prepare in dark room or place a box over the electrophoresis apparatus during gelation and electrophoresis Expired contaminated or degraded Verify that the DNA dye has DNA dye not d
6. G GenoSensor Corporation eee EduPrimer VNTR DNA Profiling Kit Catalog 3003 Version A July 2014 User Manual EduPrimer VNTR DNA Profiling Kit Manual Table of Contents Notes for INStrucCtoOlrs cccccceesseeeeeeseeeeeeseeenseenseeseeuseenseenseenseeneeeeeeneeenes 2 Shipping Storage and Safety cccccceeceseeeeeeeeeeeeeeeeeeeeeeeeeeneneeeeeeeeeeaees 3 SUGGESTIONS for Teachers wvssecnniccassnnennccnnnenasneinncnadtdennnadttnnnensnesnenancsnennantons 4 EduPrimer VNTR DNA Profiling Kit Overview ccccsesssseeeseeeseeeeeees 5 Kit Components and Storage Conditions cccccccccssssssceceeecesseseaeeeeeesceeseseuaecesecsseeseaeaeeeesesseessseaeeeeeess 5 Additional Required Materials cccccccssssssscccecscsssesssaesececesceseeeaeeeeecesseseaaeseseeecssseseaaeaeeeesessseseaaeaeeeesens 5 Introduction Theory and Background ccceeseeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeees 6 EduPrimer VNTR DNA Profiling Kit Protocol ccccssssseeeeseeeeeseeeees 7 PRE Dalat O g AAE E E E E A E E E EE A E E E E E dteeewoeseeceteneneetves 7 DNA PreparatiOM saccsivtsacceetcsacccavinnccactasucccant tace setedaccuentaccn celseaucccaveauied de aede aE aE aE dE SA ES daedra aE Eid 7 PER REICHO aeaa a stn cove a a a 8 Agarose Gel Electrophoresis oasis eioi eneee e aa Eeee Ea aee oi ob Aae dened eE AEE aiaee 9 R sults ANG DisSCUSS OMes tis e raie e aaa a ee ea e araa asaan aad raas 10 Data Analysis and C
7. V for 20 minutes and stop before loading dye has run off gel Depending on the DNA dye used caution may need to be taken to reduce exposure of gel to light Visualize under UV and record the results manually or by photography Compare individual experimental bands to positive control DNA Example of Gel Setup and Loading Run 3 gels with 12 wells each to accommodate 24 students and 6 control reactions EduPrimer DNA Profiling Kit GenoSensor Corp Lane 1 10 ul DNA Ladder Lane 2 10 ul Negative Control Lane 3 10 ul Positive Control Lanes 4 11 Up to 20 ul of each student sample Results and Discussion Mass ng Kilobases Compare your results to your fellow classmates Is this In what ways is this information useful 2 Log DNA Ladder visualized by ethidium bromide staining ona 1 0 TBE agarose gel Mass values are for 1 nna lano Fig 1 10K DNA Ladder DNA Ladder reference for band size comparison EduPrimer DNA Profiling Kit GenoSensor Corp your band pattern different from others What causes 10 Data Analysis and Class Exercise Class Allele Frequency The results from the whole class can be analyzed to determine the allele frequency within the class Record the results in the table below Observed Class Genotypes oe ng of Genotypes of individuals alleles frequency homozygous w of alleles__ Alu Total amp alleles
8. ame with the isolation of DNA polymerase from a thermophilic bacterium known as Thermus aquaticus This bacterial species lives in high temperature steam vents and therefore its DNA polymerase evolved to withstand extremely high temperatures During PCR DNA is synthesized and multiplies by 2 each cycle thus the growth of DNA copy over the reaction is exponential In theory after 30 cycles there will be 2 This is over a billion copies of DNA Yielding this amount of DNA allows the possibility of visualization through a variety of means One of the most popular visualization methods is agarose gel electrophoresis Genes and DNA The human genome contains 23 pairs of chromosomes that contain a total of thirty to fifty thousand genes most of which generally code for proteins However those genes only comprise about 5 of the genome leaving 95 as so called non coding DNA This noncoding DNA is found not only between but within genes splitting them into segments In eukaryotes non coding DNA found within genes is known as introns The sequences that do code for proteins are called exons In eukaryotes genomic DNA is transcribed into RNA molecules containing both introns and exons for a particular gene While the RNA is still in the nucleus before being transported out of the nucleus the introns in stay within the nucleus must be removed from the RNA while the exons ex exit the nucleus are spliced together to form the complete coding
9. e cycling procedure involves heating the sample to 94 C At this high temperature the template strands separate This is called the denaturation step The thermal cycler then rapidly cools to 60 C to allow the primers to anneal to the separated template strands This is called the annealing step The two original template strands may reanneal to each other or compete with the primers for the primers complementary binding sites However the primers are added in excess such that the primers actually out compete the original DNA strands for the primers complementary binding sites Lastly the thermal cycler heats the sample to 72 C for Taq DNA polymerase to extend the primers and make complete copies of each template DNA strand This is called the extension step Taq polymerase works most efficiently at this temperature Two new copies of EduPrimer DNA Profiling Kit GenoSensor Corp 14 each complementary strand are created There are now two sets of double stranded DNA dsDNA These two sets of dsDNA can now be used for another cycle and subsequent strand synthesis At this stage a complete temperature cycle thermal cycle has been completed Each step takes about 30 seconds to 1 minute and this process continues for roughly 30 40 cycles depending on how the user has programmed the thermal cycler Each step is repeated in that order each cycle until it is completed At the end the product is put on hold ata low temperature generally 4
10. egraded in storage been contaminated or expired Non specific Premature Taq polymerase Mix solutions on ice place rxn amplification replication directly to 94 thermal cycler product Primer annealing temperature too Raise annealing temperature low in 22 increments Insufficient mixing of reaction solution Mix solutions thoroughly before beginning the reaction Exogenous DNA contamination Wear gloves Use dedicated area for sample preparation Use non aerosol tips EduPrimer DNA Profiling Kit GenoSensor Corp 17 Technical Service For more information or technical assistance please call write fax or email GenoSensor Corporation 4665 S Ash Avenue Suite G 18 Tempe Arizona 85282 Tel 1 480 598 5378 Fax 1 480 755 3319 Email tech_service genosensorcorp com Web www genosensorcorp com Limited Warranty GenoSensor is committed to providing our customers with high quality goods and services Our goal is to ensure that every customer is 100 satisfied with our products and our service If you should have any questions or concerns about a GenoSensor product or service please contact our Technical Service at tech_service genosensorcorp com GenoSensor warrants that all of its products will perform according to the specifications stated on the certificate of analysis This warranty limits GenoSensor Corporation s liability only to the cost of the product No warranty is granted for product
11. ffer DNA nucleotides dNTP s of each adenine guanine thymine and cytosine DNA polymerase 2 DNA oligonucleotide primers Template DNA starting material PCR Makes Use of Two Basic Processes in Molecular Genetics 1 Complementary DNA strand hybridization For DNA to be amplified one must have a known sequence which flanks the gene of interest upstream and downstream These sequences are used to create what are known as EduPrimer DNA Profiling Kit GenoSensor Corp 12 oligonucleotide primers meaning a short 20 base pair nucleotide sequence which is used as a Starting point for DNA replication The primers are said to be complementary to their target regions so they will anneal attach to those regions specifically Primers are required by DNA polymerase because it cannot add nucleotides without a preexisting chain Complementary Strand Hybridization occurs when two different oligonucleotide primers anneal to each of their respective complementary base pair sequences on the template They are designed specifically to anneal at opposite ends of opposite strands of the specific sequence of DNA that is desired to be amplified 2 DNA strand synthesis via DNA polymerase In a PCR a special type of DNA polymerase is used that is able to withstand the temperature fluctuations required for thermal cycling Most DNA polymerases cannot tolerate the high temperatures and fluctuations from 60 C 94 C The breakthrough in PCR c
12. for the diagnosis of disease in humans or animals Do not use internally or externally in humans or animals Consider all chemicals as potentially hazardous Only persons trained in laboratory techniques and familiar with the principles of good laboratory practice should handle these products Wear suitable protective clothing such as laboratory overalls safety glasses and gloves Exercise caution to avoid contact with skin or eyes if contact should occur wash immediately with water Material Safety Data Sheet for products is available upon request EduPrimer DNA Profiling Kit GenoSensor Corp 3 Suggestions for Teachers This kit is technical in nature and an excellent tool for teaching techniques and has real links to DNA Forensics Students will enjoy extracting their own DNA and seeing their individual band separation Should you want to there are other ways to spice things up even more Here are a few possible suggestions Where s Waldo 1 Determine your own band pattern by running a gel prior to class or from a previous year 2 Either post a picture or use the ladder to draw the bands locations on the board 3 Have the students know where they are in the gel by numbers 4 Make a cheat sheet with where the students are and where Waldo is 4 Day of experiment run your sample on the gel somewhere in the middle 5 Let the students work in groups to analyze their band and find Waldo Smooth Criminal 1 Secretly pick two st
13. lass EXxerciSe ccccccccseseeeeeeeeeeeeeeeeeeeeeeeeeneeeeeeeees 11 EduPrimer DNA Profiling Kit Background ccccseesseeeeeeeeeeneeeees 12 Troubleshooting sssssssnnnnnnnunnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn nnmnnn nnmnnn nnne 17 Technical Service asssnsunsnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn mnnn 18 Literature Citation When describing a procedure for publication using these products we would appreciate that you refer to them as the EduPrimer VNTR DNA Profiling Kit Trademarks EduPrimer is a trademark of GenoSensor EduPrimer DNA Profiling Kit GenoSensor Corp 1 Notes for Instructors Kit Components and Storage Conditions Component Storage Solution A Room temp Solution B Room temp Cotton Swabs Room temp 2X PCR Master Mix 20 C Positive Control DNA Heterozygous 20 C Negative Control DNase RNase free H20 20 C DNA ladder 20 C Preparation for DNA isolation and PCR Set heat block or water bath to 95 C Instruct the students in cheek cell collection step to make sure proper technique and timing is used to ensure sufficient collection for DNA isolation Thaw 2x PCR Master Mix on ice Before opening tube spin 10 sec at 6 000 rpm or greater in a microcentrifuge Vortex 10 seconds then spin again for 10 seconds Aliquot the 2X PCR Master Mix as necessary after doing the above preparation Each package contains e
14. nough PCR 2X Master Mix for 30 reactions Use 10 ul of 2X PCR Master Mix with 10 ul template for a final PCR volume of 20 ul Up to 3 positive control materials are available for PCR reactions 10 ul from Positive Control tube 10 ul 2X PCR Master Mix for one reaction Run 1 or up to 3 negative control reactions Electrophoresis Electrophoresis reagents are not provided in the kit Please refer to the required materials list Best results are obtained by adding DNA dye i e Gel Red or Sybr Safe to molten agarose Avoid exposing the agarose gel to light It is best to store and run the gel in a dark room or cover the gel with a box during gel polymerization and the whole electrophoresis process There is enough DNA ladder to load 3 lanes with 10 ul After PCR load 10 ul of positive control reaction for up to 6 lanes After PCR load 10 ul of negative control reaction for up to 6 lanes After PCR load as much as 20 ul of student PCR reaction into a lane EduPrimer DNA Profiling Kit GenoSensor Corp Shipping Storage and Safety Shipping and Storage EduPrimer VNTR DNA Profiling kits are shipped on blue ice Components should be stored at temperatures shown in the above table At proper storage conditions components are stable for 1 year from the date received Expiration dates are also noted on product labels Safety Warnings and Precautions This product is intended for research use only It is not recommended or intended
15. nts into the solution in the tube Place sample in heat block to incubate at 95 C for 5 minutes Immediately place tube in ice until ready for next step This process continues to destroy proteins particularly those that damage DNA Load sample into microcentrifuge mini centrifuge works as well taking care that there is another sample directly across from your sample to keep the centrifuge in balance as it spins Close internal and external centrifuge lids Spin briefly 10 seconds to pool condensation that has collected on the cap Remove swab with tweezers tweezers should be rinsed with ethanol between samples to prevent contamination Add 20 ul Solution B green label to the sample tube So ution B neutralizes the harsh conditions needed for lysis preparing the solution for DNA isolation and PCR to follow Close the tube lid tightly and vortex to mix for at least 10 seconds 12 Load sample into microcentrifuge with the tube hinge pointing out balancing out your sample tube and closing the lids Spin sample for 1 minute at 12 000 rpm Look for a small clear round pellet near the bottom of the tube under the hinge This pellet contains cellular debris The aqueous solution Supernatant that has not precipitated into the pellet contains cellular DNA PCR Reaction Wear gloves and handle solutions carefully 1 2 Prepare and label a small PCR tube with your name Label both the top and side of the PCR tube t
16. o ensure clarity Ensure that the 2x PCR Master Mix is on ice and has been spun at 6 000 RPM or greater in a microcentrifuge for 10 seconds vortexed for 10 seconds then spun again for 10 seconds before opening the Master Mix tube as directed by instructor Label and prepare PCR tubes for controls Follow the table below Add 10 ul of 2X PCR Master Mix and 10 ul of supernatant from step 14 above avoid the pellet to the labeled PCR tube for a total of 20 ul as indicated in the table below The supernatant contains your genomic DNA which will become the template for polymerase chain reaction PCR Note It is preferred that the PCR reaction mix preparation is done on ice Mix the 20 ul PCR reaction mixture by pipetting in and out with the pipette and the close the lid tightly Store the sample on ice until it is ready to be loaded into the thermal cycler PCR reaction mixture tables Student Sample PCR Mixture 2x PCR Master Mix 10 ul Genomic Template Supernatant 10 ul Volume total 20 ul Control Samples PCR Mixture Positive Control Negative control 2x PCR Master Mix 10 ul 2x PCR Master Mix 10 ul Positive control DNA 10 ul Negative Control 10 ul Volume total 20 ul Volume total 20 ul Positive and negative controls give guidelines or boundaries to the experimental results The positive control will show a result for an individual with widely spaced bands near the upper and lowe
17. om person to person demonstrating the basis for the process of creating DNA profiles that are used to differentiate one person from another After completing this experiment one should be able to proficiently perform PCR and understand the concepts behind it Kit Components and Storage Conditions for a lab of 24 students Component Amount 30 rxn s Storage Solution A 6 mL Room temp Solution B 0 6 mL Room temp Cotton Swabs 28 Room temp 2X PCR Master Mix 300 uL 20 C Positive Control DNA i Heterozygous 30 uL 20 C Negative Control 30 uL 20 C DNA ladder 30 uL 20 C Additional Required Materials e Thermal Cycler Heat Block or heat plate Beaker with de ionized water water bath Tube floater Thermometer Microcentrifuge Microcentrifuge tubes Vortex Micropipettes p10 p200 p1000 Pipette tips PCR tubes Tube Racks Ethanol or ethanol wipes Electrophoresis equipment Electrophoresis supplies agarose TBE DNA loading buffer running buffer gel dye eg Sybr safe Gel Red Scissors and tweezers e UV light box or Gel Doc equipment and program EduPrimer DNA Profiling Kit GenoSensor Corp 5 Introduction Theory and Background Every one of us is different This is true on many different levels but it s an interesting point down on the genetic level In fact a large portion of our genetic makeup from one person to the next might differ by as little as
18. r limits of this VNTR region Negative control show the result if no DNA is present for the PCR DNA ladder is a standard which is used like a ruler to show how DNA of a certain length will migrate through a gel during electrophoresis EduPrimer DNA Profiling Kit GenoSensor Corp 8 PCR Parameters Program your thermal cycler as follows 1 oar wh 94 C 2 minutes 94 C denaturing 20 seconds 65 C annealing 20 seconds repeat steps 2 3 amp 4 for 40 cycles 72 C extension 30 seconds 72 C 5 minutes 4 C finished hold Agarose Gel Electrophoresis General Procedure detailed directions as given by instructor Prepare 0 8 1 agarose Set up electrophoresis apparatus and pour the 1 molten agarose for gelation For staining use a DNA dye which is added directly to the molten agarose For light sensitive dyes keep the gel in the dark during gelation either by performing in a dark room or placing a box over the gel Use at least 10 uL of PCR product to visualize results by electrophoresis on agarose gel If gel well volume will accommodate more than 10 ul a higher volume is preferred Loading dye is added to the sample to ensure that the sample will sink to the bottom of the well and properly enter the agarose gel Mix sample with loading dye according to instructor directions before carefully pipetting into a well in the gel Be sure to record which well holds your sample Run at 100
19. reaction or PCR He earned the Nobel Prize in Chemistry in 1993 due to his innovation His technique enabled researchers besides a few expert microbiologists to amplify DNA Before that amplification of DNA was extremely difficult and time consuming Now scientists in any field can incorporate molecular biology into their research with PCR Currently PCR is used in a wide variety of areas for example gene mapping DNA sequencing gene expression gene detection forensics criminal investigation medical diagnostics and genome sequencing Very few of these applications were practically possible before PCR The experiment does require an initial investment in specially made machinery but with the proper equipment nearly anyone can perform a successful PCR experiment without significant cost PCR and Biotechnology Revolutionizes an Entire Research Community PCR is capable of producing large amounts of targeted DNA from an extremely small amount of starting material Known as your template DNA can be obtained from any cell such as blood cells hair cells cheek cells etc and after proper treatment to isolate DNA PCR can be applied to create millions of copies of virtually any desired DNA sequence That is one of the most significant powers of PCR specificity Although you may put an entire genome worth of DNA into a PCR it amplifies the exact piece of DNA desired and leaves the rest out The basic components of PCR Reaction Bu
20. s beyond their listed expiration date No warranty is applicable unless all product components are stored in accordance with instructions GenoSensor reserves the right to select the method s used to analyze a product unless GenoSensor agrees to a specified method in writing prior to acceptance of the order GenoSensor makes every effort to ensure the accuracy of its publications but realizes that the occasional typographical or other error is inevitable Therefore GenoSensor makes no warranty of any kind regarding the contents of any publications or documentation If you discover an error in any of our publications please report it to our Technical Service GenoSensor assumes no responsibility or liability for any special incidental indirect or consequential loss or damage whatsoever The above limited warranty is sole and exclusive No other warranty is made whether expressed or implied including any warranty of merchantability or fitness for a particular purpose EduPrimer DNA Profiling Kit GenoSensor Corp 18
21. sequence which will soon be translated into the protein This process is called RNA splicing Some genes may contain a few introns others may contain dozens Interestingly it is the non coding junk DNA that is useful to us when considering a DNA profile of an individual instead of the DNA that actually codes for life As discussed functional segments of genes exons code for proteins Proteins are molecules that carry out most cellular functions Exon sequences are therefore very similar among individuals because even slight difference can change the function of the protein in a EduPrimer DNA Profiling Kit GenoSensor Corp 13 potentially harmful way many diseases are caused by mutated proteins Introns however often vary in size and number among individuals Intron sequences are thought to be the result of the differential accumulation of mutations throughout evolution that are silently passed to descendants through the hereditary code It is this difference in intron sequences that allows us to determine human genetic diversity The identification of these distinctive characteristics in DNA represents the molecular basis for human identification and population genetics Throughout evolution intron sequences have been the target of random insertions by short repetitive interspersed elements also known as SINEs SINEs have become randomly inserted within our introns over millions of years One such repetitive element is called the
22. udents to be the criminal This can be done by collecting all of the student supernatant DNA samples after students are done with them and selecting two of the tubes from the whole class Make note of which student s samples you selected Alternatively collect swabs from the criminal students cheek cells ahead of the class period and have them prepared alongside the rest of the class 2 Run the criminal samples through the PCR process alongside the rest of the class 3 Run the criminal samples on the same gel or gels as the rest of the class for easy comparison Each PCR reaction can be split into two gels with a 20 pl PCR product split in half into two gels This will only work if the student can keep a secret and is not the obvious choice 4 Have the students compare bands and explain the effect of Gel smiling 5 Optionally offer a simple reward to whomever figures out who the criminals are as well as to participating criminal students EduPrimer DNA Profiling Kit GenoSensor Corp 4 EduPrimer VNTR DNA Profiling Kit Overview The EduPrimer VNTR DNA Profiling Kit introduces Polymerase Chain Reaction PCR techniques to students or anyone wishing to learn PCR and its uses It contains all reagents necessary for DNA isolation and PCR PCR is an extremely important and valuable skill to have in contemporary biological and related sciences In this particular kit the experiment generates varying results fr
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