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realshinjiikari · 6 months

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Professor Snape x Teletubbies erotica. Do not open.

Severus Snape, Professor and Lover

Severus Snape was very sad. He was not having many pleasures doing lectues and teaching stuents anymore. All those adventures happening at Hogwarts sucked the inner life from his bones…and he had nothing more to accomplish. He went to Dumbledor:

"Heaadmastet, I want to be expelled, I don’t have what it takes to teach anymoire."

Dumbledore was puzzled:

"But Snepa, you are my best professor, you can’t go and waste the knowlegde you have!"

"Oh Dumbledore…I’m so sad." said Snape putting the back of his hand on his forehead.

Dumbledor, the powerful and wiseful magus he wwas thought a solution:

"Okay Snape, I’ll transfer you to another place, somewhere calm where you will help good creatures that have n o parents."

Snape got greatful thatDumblerdo understood his feelings and pushed him to place less onerous to his advancing age.

Snape took a train and after days, finally reached an colorful and wonderful place. It was the Tubbydome Supertronic, the place of dwee]lling of his new students. He reached there, and a periscophe welcomed him:

"Profwssor Snape, you arrived! The students waits you in the classroom."

Snape was still sad, but his powerful stomach got chills, because the mystery of novelty aproacches his comfort zone.

When Snape entered the classroom, four color creatues were there seatting on the chairs. They said to Snape:

"Eh-Oh!" and Snape smilled, feeling his heart get full of ingnominous passion.

"Alright dear students, today, you will learn potion making."

"No, we want to learn how to be adult!" cried the four Teletubbies. They had live for many long inside that perfect bubble, only experimenting the baby joys of youth. But every day was equal to them, not really happened at their house and life was boring.

When Snape arrive, a flash of hope filled their hearts with the promisse of life beyond the perfect prison they got trapped whne they got born. Only that dark robbed and pale skinned man can save the babies from this terrible fate.

"But kids," said Seevrus "I must acomplish the curriculum!"

"Oh professor, I’m dying from inside, don’t let the evil periscophe suck the last remnants of humanity locked in my so fatal brokened heart" saaid the yellow Teletubbie, Laa-Laa, with a sensive voice, calling for a strong man to free her mind…her body.

Snape looked her, and felt somethnig funny inside his pelvis. It was a long time since the last time he shared a intimacy filled momnet with a woman. Snape was starting to break:

"But Teletubbies…if the periscophe woman watches us…how can we have privacy?"

The purple leader, Tinky Winky, got up and uttered with his powerful throat:

"I have a secret place professor. We can learn there and free ouselves from the bounds of madness that others inflicted upon us!" said him hitting his chest with a clench hands.

Snpae looked down, thought, and followed the purple leader. They opened a secret door inside their house, a door leading to the basement.

The place was full of rats and cockroaches. Water infiltrated the place and mold smell was present in dangerous quantities. A lot of earth and dirty was covering the place, and a hole in the wall could be seen.

"You are making a hole?"

"Yes Snape," said Tinky Winky, "we must escape, so we are creating a exit route to get out of the evil crunchs of the woman periscophe."

"Oh, I must help you Teletubbies."

"Prodfessor" said Laa-Laa, "first, teach us how to adult!"

Snape nooded in agreement. He, wth his wondrous magic wand created a giant bed, and some wine and cheese to acompany this iluustrious wisdom moment.

"Teletubbies, lay her with me, I’ll teach you how to be a man. Oh, but you Poo, aren’t ready yet. You are too young baby. Stay here in the corner and play with you small hose." and Poo got sad, but neverthless, he did what Snpe said and watched the others while they got teached.

"So" said Snap, "Laa-Laa, you make a pair with me. Tinky-Winky, you go with Dipsy. Watch me while I play with Laa-Laa and mimic my movements. This way you will learn."

Snape them got on his kness and looked at the layied Laa-Laa. She was sweatting a bit.

"Don’t be afraid small yellow girl, just spread your legs." and she did. An engorged vagina shine at her crotch, but it was full of cotton yellow pubic hair. Snape picked up a scissor and cut it.

"Snape" said Dipsy "Tinky Winky have no crotch mouth, but a very strange tongue. It’s filled with purple wrinkled skin!"

"Oh Dispy, he don’t have crotch mouth. But behind him, you will find a pork’s eye. Don’t be afraid, it’s dirty, but after a while you will like the fine flavor of melted chocolate covering your lips."

Dipsy turned Tinky Winky, and the purple Telletubbie revealed his deep purple anal hole.

"What I do?" asked Dipsy.

"The same I will do with Laa-Laa, don’t be afriad. Tinky Winky will scream and moan, but you must go on. " said Snape with a calm and softfull deep voice.

"Oh professor" said Laa-Laa with a moaning lustful voise, "I think I peed myself. I’m feeling moist."

"No Laa-Laa, you not pee. You only are happy that I’m here."

"Seriously?"

"Yes. Now…suck my index finger" and she did.

And Snape unclothed himself. His pale shiny penis appeared and all of the Teletubbies got impressed. His nut sack was very white and hairy and exhalled a snake oil parfum essence. Laa-Laa felt a jolt of pleusre down her antenna.

"Now boys, do as I do" and them, Snape introducted his wizard wondrous penis inside Laa-Laa. She screamed:

"AAAAAAHHHHHHHHH!" and a flush of yellow blood got expelled from her now broken hymen. The yeloow blood covered Snake, and he rubbed himself with that sticky liquid.

"Prodessor" moaned loudly Laa-Laa, "isn’t this wrong? I’m feeling so dirty."

"No Laa-Laa, don’t be ashamed. When you bit the forbidden fruit, the knowlegde will fullfil you inner most desires. Be my Eve!" and Snape punch her uterus with his roquefort penis. The Teletubbie girl moans more and smilles her face.

Dipsy do the same on Tinky Wonky, puncturing the purple asshole with the green penis he is so proud of. Tinky Winky screams:

"AAAAHHHH! You hurts me my dear friend! Stop, I cannot withstand your protuberant obelisk of delectation!"

"I can’t" said Dipsy with a condensending voice "professor said I must go on with our journey throughout the steps towards a greater understanding about our true nature as alienated species from our parenthood."

"Tinky Winky was suffering the destiny of those under the yoke of their own expectations. But Snape was wise and appealed:

"Dipsy, use your bodly fluids to appease the sorrow of your roost companion."

Dipsy them spit on his hand an immense gob of putrid mucus. It was green as his body and full of dancing crawlers that were enjoying the hot abode that was his nostrils. Dipsy covered his fluffy hard penis with it and penetrated Tinky Winky again. This time, the purple one enjoyied.

"Ooohhohoohohhhhoohhhhh…"

Snape was funcking hard Laa-Laa. A lot of yellow blood continued to get ot her defilled vaginal daisy. The pain hurted her much:

"Profezro, It aches my skin and negates the maturation you unleash inside my sacred womb."

"Be calm my yelloew student. After a while, no pain can touch you. Wait the massage of gods." and she did. After a while, the orgasms started, and she had multiple ones. Her chest grew a pair of xanthous brests, and they lactated the nectar of the houses of holy.

But she wasn’t in her mature years yet, and her bowel controls were rudimentary. The orgasms unleashed a cataract of pappy hot shit thru her untouched lutelous anus. The slop of dung covered the bed and the smell was atroucious. The flow was so intense, globs of shit splashed at Snape’s legs and hit his pale and sweet face.

"Ohhhhhh Laa-Laa" said Snape "finally, the cocoa honey bestow us with this feast of gormandized gut’s contents!" and Snape rubbed his face with the black fetid hodge-podge ooze Laa-Laa vomited with her anus. It so was hot and disgusting that Snape vomited a mash of orange juice, bloody spaghetti and mama’s chilli pepper over her lustful body. It reached Laa-Laa vagina and burned it and splashed all over her tummy. She rubbed the vomit all over her belly and breasts and swallowed a good portion of it.

Laa-Laa is happy, she reached a new stage of illumination and now can let go her shame of being the mistress of a snake old man.

Dipsy took a portion of that hot boiling dung and deposited it inside Tinky-Winky’s anus. Now, the penetration is complete. They are enjoying the most complete recollection of human’s past lives as beings dipping inside their own bodly properties.

"Snape" said Laa-Laa, "can I penetrate you too?"

Snape got pensive. He never thought about something so insulting to his manhood, but the pleasure was so insane, he nooded with a moan:

"Yeeeeesssssss" and Laa-Laa prepared her finger full of vomit and macaroni strings. She moved slowly her index finger and prepared to penetrated it…and she did! The finger entered Snape left nostril and started to fuck it. She was fast and didn’t want to lose time.

As Laa-Laa fucked Snape’s nose, the vomit in her nails entered it and he breathed it. He gaged and coughed the bloody puke and it hit Laa-Laa’s eyes, but she laughed.

Laa-Laa them put her other index finger inside the other Snape nostril, and he felt the pleasure more intense. Laa-Laa’s tits lactated like no other in the world, like a fountain of milk and passionate dung. The milk and shit shower was so intense it reached Poo, that was at a corner masturbating to the whole scene. He drink the shitty covered milk and peed himself in pleasure. Even a small Telletubie have the right to fell the magnificent joy of the motherly nourishment coming from his sister’s breasts.

Now, Snape is ready to cum. He grabs his penis and fell the throobing coming…and them…the cum-tastic delight goes out of his powerful snake hose and fills Laa-Laa’s small vagina with a gorgeous and thunderstriking goo that overflows her and wash the last pureness that yellow and sexy body had one day.

She cums too, and now, she is a complete fl, a yelow and tasty alien woman girl with nice and softly cotton breasts. Her Telletubbie body got gorugeously tasty an even her brothers think she is delicious now that she have breasts.

Dipsy cums inside Tinky Winky too, and the two cotton alien boys now discover what it takes to make a real action hero.

"Boys" said Snape "you have graduated. You now are full fledged mature and responsible adults. You can take care of your own butts and do with it all you deserve."

Poo was a little sad, but she know her time will come.

"Snape" said Tinky Winky rubbing his cum covered and hurting ass "We must escape, the perischope woman is already searching for us, because we haven’t returned to sleep."

"Okay Tinky-Winky, you lead the way" and the Telletubbies and Snape followeed Tinky-Winky’s down the hole they excavated. The cave was very dark and long, but Snape used his wand to iluminate the way.

As they walked, down the cave, a light at the end of the tunnel was near, and they raced towards it. But someone was expecting them. A figure was facing the cave, with his back turned against the light out of the tunnel.

"You shall not pass." said the voice

"Who are you?" askjed Snape

"I’m very disappointed with you Snape. I thought you had a rocky shadow heart. But it seems I’m wrong. It was a mistake sending you here to placate these little rainbow bastards. A mistake I’m going to correct."

"Wait, I know that voice…it’s-" and them, the figure emerges from the light, create a flowing flame around him, and his face is revealed. It’s Dumbledore!

"No! You!" shouted Snpae.

"Yes, me!"

"Why do you want with the Telletubbies?"

"Oh, you don’t know? They are Death Eaters Snape! We must kill them!"

"Death Eaters? That’s impossible!"

"No…" said Tinky-Winky "it’s true. We worked to Voldermot."

Snape got awed:

"No…why?"

"Voldermort promissed that he would free our homeland from the dementors horde invasion. But, as we worked for him and infiltrated inside the Hogwarts, Dumbledore caught us. Now he absorbs our energies to fuel his mecha-bulldozer that-"

"Shut the fuck up!" cried Dumbledore. "You now will die!"

"No, I will not permit!" said Snape. He created a strong bound with those aliens, and he can’t let them die, even if they are working for the enemy.

"So" said Dumbledor "you will die too!" and Dumbledore invoke the killing speel from his wand. But them, Yinky-Winky, from the bottom of his purse, pulled out a Colt 45 Revolver and shoot Dumbledore, but the wizard deflected the bullet.

"HA, you cannot kill me!"

"Snape, go, save the Telletubbies, I’ll buy you time!" cried Tikny-Winky.

"No!" shout Laa-Laa "we can’t let you die here!"

"But you must, our he will kill us all and no one will can save our homeland!" and the Tinky-Winky advanced in the Dumbledore’s direction shooting rapid bullets at him.

Snape used his wand and started to drill a hole in the cave, circumventing the vicious battle happening near the entrance. Snape and the Telletubbies managed to drill another hole out some feet awya from the confrontation and gotout of the cave. They raced away from it, and at some distance, they saw Tinky-Winky shooting Dumbledore:

"Dubledor!" cried Tinky-Wink. He picked a hand grenade, pulled the pin, threw it at Dumbledore and shout "Die BASTAAAAAAAARD!"

"NOOOOOOOOOO!" and the cave exploded, lauching debris all over the place.

"Fast Telletubbies," said Snape "HIDE!" and they did. A lot of rock felt from the sky and fumes and dust covered everywhere.

"Tinky-Winkyyyyyyyyyyyyyy!" cried the three Telletubbies, and Laa-Laa tried to run and find her purple brother, but Snape prevented:

"No, you can’t! This place is unstable, we will die staying here!" cried Snape and the earth started to shake. They raced away from the crumbling Tubbydrome Supertronic, avoinding the falling rocks and the posion flower that attacked them.

After a while, they reached a safe haven.

Laa-Laa was very sad and crying much:

"Why? Why we let him die Snape?"

"Oh Laa-Laa, he saved us and was his desire that we escaped."

"But it is not fair…"

"I know…I know" and Snape hugged Laa-Laa, and the other Telletubbies did it too.

Now, these four heroes must find a way to save the Telletubbie planet from the horde of dementors that are trying to invade and conquer the place, slaving all of the millions of telletubbie inhabitants. Who is behind all this? Is Voldermort involved? And Snape and Laa-Laa? Will they be together till the end?

Snape says:

"Laa-Laa, I promisse, we will avenge your brother…"

The End?

#this has been living in drafts for yeara #time to set this little dickens free

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chloeafrazier · 21 days

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My View On Severus Snape From The Harry Potter Movies.

Hey Everyone how are you all doing today ? Today i would like to talk about Harry Potter so i have been watching the Movies Like crazy

Lately and i would like to take the time to defend one of my Favorite Character's in the entire series and that is Severus Snape Now i know

people have preconceived notion's about snape and Look i am going to be very truthful for a Long Time I did not Like snape for a Long

time it took me years and years to like him as a Person and and a Character look i know that snape is a very Controversial Character i

personally was not his Biggest Fan for a long time but after watching all of the Moves and doing research on his Character online and all i

have found that he was a Person who was place by Dumbledore and a few others now i do not Blame Dumbledore but people gave snape

a hard time about him killing Dumbledore but you have to Understand that Dumbledore Told Him to do it if you Remember In

The Third to the Last One of the series In Harry Potter and the Half

Blood Prince He tells Dumbledore that he is tired of being a Double agent he says has it Ever Crossed That brilliant mind of yours that i

might not want to do this anymore ? and in the Deathly Hallows Part 2 Dumbledore says You Must be the one to kill me Severus Only Then

will the Dark Lord Trust You Completely so if you stop to think about it in a way Dumbledore did not really Give Severus a Choice but to kill

him and that i feel had to be very hard for snape Because you could tell by the Look on his face that he did not want to do it for sure at

least that is the way i see it in a way when it was all said and done as far snape killing Dumbledore that is in a way it was not like he Gave

him much of a Choice that is for sure Look i am saying that agree with Everything that snpe did okay but what i am saying is when it is

all said and done it was Not like he was Given Much of a choice that is for sure and that is My View On Severus snape From The Harry Potter Movies well Thanks for reading have a Great day.

#A New Blog Post About Snape From Harry Potter.

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zehub · 2 years

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#Publicités #lumineuses: l'#extinction #nocturne confirmée par le Conseil d'Etat

Le Conseil d'Etat a rejeté mercredi la requête en référé-suspension formulée par le Syndicat national de la publicité extérieure (SNPE) contre un décret qui généralise l'extinction des publicités lumineuses pendant la majeure partie de la nuit, selon une ordonnance consultée par l'AFP.

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juotte · 2 years

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snpe 지난달 판매량 집계 상위 10개 추천! 장점/단점은?

snpe 판매량 상위 10개를 추천해드리겠습니다~ 추천 순위는 네이버 쇼핑 트랜드, 쿠팡 판매 순위, 구글 키워드 트랜드 등 주요 온라인 쇼핑몰의 판매 데이타를 분석해서지난달 가장 많이 판매된 snpe 제품상위 10가지를 집계/정리해서 알려드리고 있습니다.~빅데이타를 기반으로 선정된 최다 판매 snpe 추천 인기 제품 상위 10개입니다.아래 보여드리는 리스트는 snpe 찾는 소비자들이 가장 좋아하고, 판매가 가장 많았던 HOT snpe 제품 중 상위 10가지 제품입니다. snpe 구매하시는데 참고가 되셨으면 합니다^^ 이 중에 골라보시면 후회하지 않으실거에요^^ 이미지를 클릭하시면, 상세한 제품 소개와 상품평 확인하실 수 있으니 실재 구매하신 분들의 상품평도 꼭꼭 확인하시는 것도 잊지 마시구요^^~~시간이…

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sips---tea · 3 years

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Caught // Severus Snape x reader one shot

"Severus?" you asked, walking into the spare potion cupboard. Severus had asked you to meet him with a certain plant he needed so you sat on the ladder waiting for your boyfriend to arrive. After a short, while the door opened to see Severus, he smiled down at you as you got up from sitting and hugged him gently. "Hey Severus" you said smiling.

"(y/n)" he said with a smile. You gave him the plant and he went up the adder gracefully and placed it on one of the high shelves.

"Careful" you said, worrying.

Severus rolled his eyes and glided down the ladder with ease and stood in front of you again. He took your cheek and you leaned into his touch, smiling. "Why do you worry so much?" he asked.

"Because I love you" you said smiling up at Severus. "And I care for your wellbeing"

Severus smiled and stroked your cheek, shaking his head gently, he found it funny how much you cared for him, even with small things that he did. He met your lips with his and he sweetly kissed you. You grabbed Severus' robes and smiled into his kiss, enjoying how gentle and loving it was. The kiss soon grew more passionate and Severus moved his hands from your cheek to your hair, running his fingers through it.

"Severus" you murmured after a while of passionate kissing.

Severus let go and raised his eyebrows.

"I have to teach soon" you murmured.

"Surely they could spare you for longer" Severus said, he took your cheek again and stroked your lip with his thumb.

"Yeah I am sure they could" you said with a smile, holding Severus's hand to your cheek. Just as you held it a figure walked in front of the cupboard and looked in. Harry stood, a confused look on his face, his eyes widened when he recognised the two teachers and quickly Severus closed the door.

"Oops" you murmured. Severus smirked and stroked your cheek and you shook your head. "I have a lesson with Harry now I need to be there in time."

Severus huffed and let go of your cheek and he hugged you into him like he never wanted to let you go. He huffed again and kissed your head before realising you.

You opened the door and blew him a kiss. "Love you Sev" you said

Severus smiled and when you closed the door caught the kiss in his hand and kissed it. He took some ingredients out and stalked down the hallway, missing you already.

#snape fanfiction #snape #severus snape #harry potter #caught #short fanfic #oneshot snpe #snape x reader #severus snape x reader #severus x reader #snape x (y/n)

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spainkitty · 4 years

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As much as I love Stiles Stilinski, I would never say he's... I dunno, unproblematic?? The whole point of loving him, for me, is that he IS flawed. He's relatable and upsetting and real. I don't agree with all his actions or what he says every time. I love seeing his character develop and CHANGE because, again, he's not a perfect, flawless person. He's breakable and human and flawed. Arguing he's a bad character because he's flawed and fucks up sometimes misses the point of... I dunno... writing a character???

When he shouts at Scott about how some people are human, I didn't feel it as a jab at Scott's "monstrosity". I felt it as a jab at Stiles' own insecurity and knowledge of his own flaws (and disbelief in his own self "unworth" lol? making up words evurrday cuz why else would I have a ling degree, amirite?). Yes, because Scott had his own doubts and insecurities, perhaps he took it differently and it hurt him in a way Stiles did NOT intend. Because, again, they are fallible people. Different things hit people differently (obviously). It showed perfectly how far apart these friends had fallen, actually. Neither could see the other's point of view.

In a very basic way: the part of the Stiles fandom that sees Stiles as an Ultimate Hufflepuff, True Pack Mom, soft boy, I don't.... really get? To me, he is, and always will be, a total Slytherin. He's clannish, ambitious(kinda?), determined and driven, and rather selfish, and very cunning. He cares about the people HE cares about and he will throw himself into the fire (or the ice water that legit kills him for... 16 hrs?) for those select important people. Everyone else can go burn. I LOVE KouriArashi's (did I remember that correctly?) Searching Ceremonies series because they capture SO WELL this dichotomy of his personality: he's a Denmaker and wants to take care of the people he loves. However, he's also capable of being the Lefthand, getting his hands dirty and making questionable choices for the sake of the ones he loves.

His "everyone else can go burn" mentality is NOT a "good" trait, but it's the leading part of who he is and it's actually kinda relatable. And, because he's a FANTASTICALLY written, grey moralled teenager played by an amazingly charismatic actor, there's a whole lot more to him, he also changes and develops throughout the show. But he is NOT a "perfect, does no wrong, and has zero flaws" character.

And this is my big beef with a certain circle of Scott fans. There's a whole lot of viciously defensive people who will go on for PAGES AND PAGES about how perfect and wonderful and flawless and amazing the Scott is. And, even worse, the writing of TW wants the watchers to believe it, too. Some fans believe this is intentional and Scott is just an unreliable narrator. Power to you. But I'm old and cynical and I hate JD, so I honestly believe it's just bad writing. He really is that kinda bland 1D character. Using the defense/argument that "My character is more perfect and morally superior than yours" is a dumb argument. Characters are supposed to be flawed, are supposed to grow and change. We're supposed to disagree or cringe through their actions and choices, so we can cheer on their visible growth!!

Scott's a bad character because he doesn't grow. Not because he does bad things.

Stiles is good character because he does grow, even if he does do bad things.

#long post #stiles stilinski #Slytherin Stiles #Every Time #i was going to say always but SNPE FUCKING RUINED IT FOR ME #twreck #unpopular opinion???#maybe??#eh whatever #anti scott mccall

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snictionary · 6 years

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snartist

/sn ɑː tɪst /

noun plural: snartists

Someone who makes snart.

“Who’s your favourite snartist?”

#snictionary #severus snpe #snapedom

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jerukjerukan · 6 years

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Me: “Wow, I think Adam Driver’s performance in TLJ is amazing and quite close with how I pictured snape acted in POA :)))”

My Snape mutuals:

#please guys i just want to live :'')#i'm thirsty and need more snape content guys #adam driver quite provide it tbh lmao #i didn't want to start a discourse man i just want to eat #this is just a joke btw :")#for some of people who agree with me i love you fam #severus snpe #snape #pro snape #jeruk talks

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jrobertsons · 6 years

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JAM 💦💦💦💦

#DAS A SNPE

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gothregulus · 3 years

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Today i came across a post about how regulus stans love him despite being a wizard nazi (and hate snpe for that reason i guess ) and i immediately thought of your post -the one that says "if regulus was real i'd beat him up" .

keep up the good work :')

ngl that post was funny af and no one makes fun of regulus more than regulus stans

thanksies !!

#asks #i want that twink obliterated

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juotte · 2 years

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snpe 지난달 판매량 집계 상위 10개 추천! 장점/단점은?

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perfectlytinyworkspace · 4 years

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DNA polymorphisms

DNA polymorphism (DNAPM) – segment of genomic DNA that differs from others in an individual

DNAPMs can be single nucleotide variations, small insertions or deletions, duplications / number of duplications in or of a segment

most are anonymous DNA polymorphisms which are not phenotypically reflected in any way

5000 out of millions of DNAPMs actually alter amino acid sequences in humans

context: less than 2% of human gDNA makes expressed codons out of 3E9 bp total haploid h-genome | 5% are coding sequences

also, codons can be spliced out and are degenerate anyway - so majority of human genome is non-expressed

still, non-expressed anonymous DNAPMs are experimentally useful – serve as markers for locating linked, co-inherited disease mutations

types of DNA polymorphisms

1. single nucleotide polymorphism (SNP) – difference in one sole base pair between individuals and populations

in a bi-allelic SNP, changed allele is the derived allele and unchanged allele is the ancestral allele | most SNPs thus show low heterozygosity

SNPs occur with low frequency and thus can reflect relative timing of a SNP group’s divergence from a common ancestor species or ancestral population

when using SNPs to order ancestral diversion events, careful of reversion derived mutation that seems to indicate a doubly-derived individual or population’s closer relation to the ancestral group

RFLPs are SNPs that specifically affect particular RE recognition sites, altering recog. sequence and losing the cut spot or creating new recog. sequence

2. deletion-insertion polymorphism (DIP) – difference in number of base pairs present

less frequent due to tendency to more heavily alter coded information in DNA sequence

often 1-2 bp only, resulting from DNA replication or recombination mistakes or breaks in DNA

in promoter and coding regions, DIPs cause frameshift mutations unless in multiples of 3 for trinucleotidic codon-based reading frames

3. simple sequence repeat (SSR) – aka microsatellite, difference number of tandem duplicates of a basic repeated sequence (≤ 10 bp)

SSR occurrence = most often silent and in non-coding areas

SSRs in coding regions have dire consequences on phenotype and are generally lethal or severely debilitating

initial creation of repeated sequence is infrequent, but once created the SSR causes DNAP to slip and generate more repeats during replicative synthesis → so, many SSRs = highly polymorphic on an individual or population level

stability + high polymorphism rate (high heterozygosity) = good marker for linked disease alleles (i.e., disease allele is linked to n repeat number at SSR | non-disease allele linked to n+a repeat number)

4. copy number variant (CNV) – difference in number of tandem duplicates of a larger repeated sequence (≥ 10 bp)

mostly stable and inherited rather than newly mutated in an individual | serves as a good pedigree marker for a linked disease allele in a family

CNVs increase possibility of unequal crossing over between homologs that results in larger or smaller number of repeats

genotype anonymous DNA polymorphisms to:

1. map regions in the physical genome and locate linked disease gene loci

» requirements for assessed individuals

different phenotypes in a single relevant gene

heterozygous individuals to assess recombination in

many individuals to test for recombination frequencies

» requirements for isolated DNAPM marker

1 DNAPM occurs every 1kb » can calculate number of DNAPMs available for mapping along gene via number of possible DNAPMs for mapping = gene length • 1 DNAPM / 1000 bp

must be unique and detectable in individual’s genome to work as a marker, as indicated by name

direct detection: if DNAPM is the disease-causing mutation, then DNAPM shows perfect linkage to disease gene

indirect detection: if DNAPM is not the disease-causing mutation but only closely linked to it, then DNAPM-disease allele recombination must be taken into account when assessing individual genotypes

» determining marker SNP original and variant allele sequences

use restriction fragment length polymorphisms (RFLPs)

RFLPs are relatively rare, but very detectable via probe bind, RE cut, and Southern Blot

added or removed site results in differing sizes of fragments hybridized to the probe

location of probe can indicate varied RE site location + reasons for why fragment lengths are or are not visualized

use allele-specific oligonucleotides (ASOs) in a DNA microarray

attach an ASO – oligonucleotide strand with specific SNP allele – to a fluorescence-detecting chip

fragment genomic DNA into probe-like pieces, amplify, and place on replica nylon membranes for each SNP allele

hybridize gDNA to ASO in high-stringency conditions on each membrane and measure degree of fluorescence to determine whether specific SNP allele is present

use single nucleotide primer extensions with specific primers and ddNTPs

less time-consuming than ASOs, so SNPE is a high-throughput technique, many samples at fast pace

sequence a primer for segment preceding location of SNP, so that primer ends on the base pair immediately before the SNP base

mix with sample in four different wells/tubes and add a different non 3′OH ddNTP to each so that primer is terminated after 1 base addition

if ddNTP complements allelic base at the SNP locus, the sample is visualized to be 1bp longer under gel electrophoresis

» detection of DNAPMs and associated disease alleles take place across Southern Blotting of family pedigrees

determine the following before labelling which DNAPM fragments are associated with mutant or WT disease alleles:

disease gene dominance, DNAPM linkage to disease gene, and individuals’ disease genotypes

more informative markers determine P(diseased), less informative markers assess P(non-diseased)

disease probabilities derived from less tightly-linked DNAPM markers must take linkage RFs into account

2. identify individuals for forensic purposes

» use SSR-allele combinations to identify specific individuals by their DNA fingerprint

probability of unique SSR repeat-number combination at multiple SSR loci = extremely small, so individual’s multi-SSR (13) genotype ≈ a DNA fingerprint

to determine individual’s multi-SSR (13) genotype, use PCR amplification of DNA sample with each SSR primer differentially fluorescent-labeled

using gel electrophoresis, assess fragment sizes for allele present at each color-distinct locus

» moral question: family members’ DNA can be utilized to identify criminals forensically, but the strategy places relatives under constant surveillance

#genetics #notes

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