A rematch from 2014 when Recs strode into round two with a 46-4 win on home turf.
Aberdeen are the reigning Scotland Rugby League champions and will be keen to make the most of home advantage and get revenge for three years ago.
Their 2015 heroics, when they lost 27-20 away to leading NCL side Skirlaugh, show they could be capable of a shock against Pilks, who finished eighth in the NCL Premier Division last season.
NCL Premier Division's Egremont breathed a sigh of relief at the end of last season as a late surge of form saw them edge away from relegation trouble.
The Cumbrians made it to round two last year and will be favourites to progress again on Saturday against the Aldershot-based side, who lost 30-4 on home turf against Siddal in round one last season.
Cumbria Men's League champions Distington were the only regional league side to make it to round two last season, but will face a stern test against the Lions on Saturday.
Fev finished fourth in NCL Division 1 last season and lost out to Skirlaugh in the second tier's promotion play-off final.
Former Wakefield Trinity star Jamie Rooney is Head Coach of Featherstone Lions
They lost out to Skirlaugh in the NCL Division 1 promotion final
They made it to round four in 2016, impressively beating League 1's South Wales Ironmen along the way.
A fierce local derby is in store between Castleford-based Fryston and Normanton, from Wakefield.
The Warriors secured their place in the 2017 Ladbrokes Challenge Cup by storming to the winter Pennine League title.
They last appeared in the Challenge Cup in 2011, when their run came to an end in round three with a 64-10 defeat at Batley Bulldogs.
Normanton were on the end of a shock as Distington produced a 31-18 win in West Yorkshire and will be keen to avoid another round one exit this year.
The BBC Sport cameras will be in attendance for the first time as Miners host their local rivals at Twist Lane.
Streamed live on the BBC Sport website, a crowd in excess of 1,000 is expected to gather as the two NCL Premier Division big guns go head-to-head.
The two sides picked up a win each against one-another last season, with just three places separating them on the league ladder.
St Pats return to the Ladbrokes Challenge Cup for the first time since 2014, when they were stopped by Leigh Centurions 74-6 in round three, while Miners were stunned 20-14 at home by Division 1 Underbank Rangers last season.
But the hosts on Saturday will be looking for a repeat of their efforts in 2015, when they thrashed League 1 Oxford a 32-6 thrashing of League 1's Oxford at Leigh Sports Village was the highlight of their run to the fifth round.
Full Time Match Official Scott Mikalauskas will be taking charge of the game.
The first of three games in the London triple-header, London Chargers won the Conference League South last season to secure their place in round one, and will be taking on Welsh champions Bridgend at London Skolars' New River Stadium.
Both sides are yet to pick up a Challenge Cup win having both suffered first round defeats on their respective debuts in the competition last season.
The Lincolnshire-based RAF welcome York Acorn on Saturday.
The Air Force lost 26-4 at Wath Brow in round one last year and lost picked up a Challenge Cup win in 2014 when they overcame Dewsbury Celtic 38-10.
Acorn, who were relegated from NCL Premier Division last year, stand in their way and will be hoping for similar success to last year's Challenge Cup journey, where they made it to round three and lost out in a memorable derby with York City Knights in front of 2.293 people.
Hull-based Myton are set for a 558-mile round-trip to Portsmouth on Saturday to take on a team who have enjoyed success in Rugby League's oldest competition in recent years.
Notably, they overcame NCL giants Leigh Miners Rangers in 2012 and ran Hunslet RLFC close in a 34-14 defeat at South Leeds Stadium later that year.
They were edged out 20-18 by Featherstone Lions in the first round last season, while Myton, who earned promotion to the NCL Premier Division last year, return to the competition after not entering in 2016, when they were beaten 34-16 by Normanton Knights in round one.
NCL champions Siddal come into this game against Leeds-based Milford as strong favourites, having claimed both the League Leaders' Shield and Grand Final titles, while Milford battled to a play-off place in Division 1.
Siddal made headlines with a shock 30-4 demolition of eventual League 1 Shield winners Newcastle Thunder in round three last season, while the Marlins' last Challenge Cup tie came in 2014 when they entertained a four-figure crowd at Leeds Rhinos' Headingley Carnegie Stadium in a 18-10 loss against then-Championship side Barrow Raiders.
Two teams who were promoted from the NCL's second tier last season square up in St Helens this weekend.
Thatto's impressive Women's team have won the Women's Challenge Cup for the past three seasons and, while their male counterparts aren't expecting to reach the same heights, they finished 12 points ahead of Skirlaugh in the NCL's second tier last season and will fancy their chances of a first Challenge Cup win since 2014.
Skirlugh, meanwhile, will be hopeful of a repeat of their last appearance in the Cup, when they reached round three, where League 1's Gloucestershire All Golds ended their run.
NCL Division Two outfit Thornhill earned their place in the Challenge Cup after winning the BARLA National Cup last season.
The Dewsbury-based side lost 8-6 at home to Hull Dockers in round one last season, while Lock Lane captured the imagination with a journey to the fifth round which included a 37-22 defeat of League 1 Oxford away from home and a 30-16 win over local rivals Featherstone Lions in front over over 1,500 people, before their run ended in round five at Halifax.
The student game's representatives this year are in for a stern test against 2016's Challenge Trophy winners.
The University outfit are one of five Hull-based clubs in the 2017 Ladbrokes Challenge Cup first round and will be hoping to avoid a second-successive first round defeat after 2016's 32-14 defeat at Millom.
Mayfield, meanwhile, enjoyed a fine run last season which ended with an entertaining 14-40 home defeat to Rochdale Hornets, who went on to earn promotion to the Championship last season.
Wath Brow wil be strong favourites for this clash having produced a fifth-placed finish in the NCL Premier Division last season.
The Hornets enjoyed a fine run to the third round last year and ran League 1 champions Toulouse Olympique close in a 14-32 defeat at the Hornet's Nest.
St Helens-based Haydock will provide the opposition this weekend after winning the North West Men's League title.
They are making their first appearance in the Cup since 2005, when they reached the third round before an admirable performance in losing 4-46 at home to Hunslet RLFC in front of almost 1,000 fans.
A huge local derby which piqued the interest of the local audience at the first round draw which was held in The Deep, Hull.
An all-Hull tie in the first round of @TheChallengeCup - West Hull against Hull Dockers!— Hull FC (@hullfcofficial) January 5, 2017
West Hull finished second in the NCL Premier Division while Dockers suffered relegation from the top tier, so the hosts will come into the game as firm favourites.
Wests came out on top in the two league clashes between the two sides last season and made it to round three in 2016, eventually halted by League 1's Doncaster in a 54-0 defeat.
Meanwhile, Dockers progressed to round two where they were beaten by NCL Division 1's Featherstone Lions, 12-14, on home turf.
Wests Warriors, who only made their competitive debut as a club in 2014, are making their Challenge Cup bow this year.
With 35 New Zealand nationals amongst their 50-man squad, they have become the official home of New Zealand Rugby League outside the southern hemisphere.
The Warriors were founded just three years ago
Wests Warriors won the Harry Jepson Trophy in September
West Yorkshire-based GB Police, meanwhile, lost out 36-4 at West Hull in round one last year and will be keen to fare better this time around.